Secreted and transmembrane polypeptides and nucleic acids encoding the same

ABSTRACT

The present invention is directed to novel polypeptides and to nucleic acid molecules encoding those polypeptides. Also provided herein are vectors and host cells comprising those nucleic acid sequences, chimeric polypeptide molecules comprising the polypeptides of the present invention fused to heterologous polypeptide sequences, antibodies which bind to the polypeptides of the present invention and to methods for producing the polypeptides of the present invention.

FIELD OF THE INVENTION

[0001] The present invention relates generally to the identification andisolation of novel DNA and to the recombinant production of novelpolypeptides.

BACKGROUND OF THE INVENTION

[0002] Extracellular proteins play important roles in, among otherthings, the formation, differentiation and maintenance of multicellularorganisms. The fate of many individual cells, e.g., proliferation,migration, differentiation, or interaction with other cells, istypically governed by information received from other cells and/or theimmediate environment. This information is often transmitted by secretedpolypeptides (for instance, mitogenic factors, survival factors,cytotoxic factors, differentiation factors, neuropeptides, and hormones)which are, in turn, received and interpreted by diverse cell receptorsor membrane-bound proteins. These secreted polypeptides or signalingmolecules normally pass through the cellular secretory pathway to reachtheir site of action in the extracellular environment.

[0003] Secreted proteins have various industrial applications, includingas pharmaceuticals, diagnostics, biosensors and bioreactors. Mostprotein drugs available at present, such as thrombolytic agents,interferons, interleukins, erythropoietins, colony stimulating factors,and various other cytokines, are secretory proteins. Their receptors,which are membrane proteins, also have potential as therapeutic ordiagnostic agents. Efforts are being undertaken by both industry andacademia to identify new, native secreted proteins. Many efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel secreted proteins. Examples ofscreening methods and techniques are described in the literature [see,for example, Klein et al., Proc. Natl. Acad. Sci. 93:7108-7113 (1996);U.S. Pat. No. 5,536,637)].

[0004] Membrane-bound proteins and receptors can play important rolesin, among other things, the formation, differentiation and maintenanceof multicellular organisms. The fate of many individual cells, e.g.,proliferation, migration, differentiation, or interaction with othercells, is typically governed by information received from other cellsand/or the immediate environment. This information is often transmittedby secreted polypeptides (for instance, mitogenic factors, survivalfactors, cytotoxic factors, differentiation factors, neuropeptides, andhormones) which are, in turn, received and interpreted by diverse cellreceptors or membrane-bound proteins. Such membrane-bound proteins andcell receptors include, but are not limited to, cytokine receptors,receptor kinases, receptor phosphatases, receptors involved in cell-cellinteractions, and cellular adhesin molecules like selectins andintegrins. For instance, transduction of signals that regulate cellgrowth and differentiation is regulated in part by phosphorylation ofvarious cellular proteins. Protein tyrosine kinases, enzymes thatcatalyze that process, can also act as growth factor receptors. Examplesinclude fibroblast growth factor receptor and nerve growth factorreceptor.

[0005] Membrane-bound proteins and receptor molecules have variousindustrial applications, including as pharmaceutical and diagnosticagents. Receptor immunoadhesins, for instance, can be employed astherapeutic agents to block receptor-ligand interactions. Themembrane-bound proteins can also be employed for screening of potentialpeptide or small molecule inhibitors of the relevant receptor/ligandinteraction.

[0006] Efforts arc being undertaken by both industry and academia toidentify new, native receptor or membrane-bound proteins. Many effortsare focused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel receptor or membrane-boundproteins.

[0007] 1. PRO196

[0008] The abbreviations “TIE” or “tie” are acronyms, which stand for“tyrosine kinase containing Ig and EGF homology domains” and were coinedto designate a new family of receptor tyrosine kinases which arc almostexclusively expressed in vascular endothelial cells and earlyhematopoietic cells, and are characterized by the presence of anEGF-like domain, and extracellular folding units stabilized byintra-chain disulfide bonds, generally referred to as “immunoglobulin(IG)-like” folds. A tyrosine kinase homologous cDNA fragment from humanleukemia cells (tie) was described by Partanen et al., Proc. Natl. Acad.Sci. USA 87, 8913-8917 (1990). The mRNA of this human “tie” receptor hasbeen detected in all human fetal and mouse embryonic tissues, and hasbeen reported to be localized in the cardiac and vascular endothelialcells. Korhonen et al., Blood 80 2548-2555 (1992); PCT ApplicationPublication No. WO 93/14124 (published Jul. 22, 1993). The rat homologof human tie, referred to as “tie-1”, was identified by Maisonpierre etal., Oncogene 8 1631-1637 (1993)). Another tie receptor, designated“tie-2” was originally identified in rats (Dumont et al., Oncogene 8,1293-1301 (1993)), while the human homolog of tie-2, referred to as“ork” was described in U.S. Pat. No. 5,447,860 (Ziegler). The murinehomolog of tie-2 was originally termed “tek.” The cloning of a mousetie-2 receptor from a brain capillary cDNA library is disclosed in PCTApplication Publication No. WO 95/13387 (published May 18, 1995). TheTIE receptors are believed to be actively involved in angiogenesis, andmay play a role in hemopoiesis as well.

[0009] The expression cloning of human TIE-2 ligands has been describedin PCT Application Publication No. WO 96/11269 (published April 18,1996) and in U.S. Pat. No. 5,521,073 (published May 28, 1996). A vectordesignated as λgt10 encoding a TIE-2 ligand named “htie-2 ligand 1” or“hTl1” has been deposited under ATCC Accession No. 75928. A plasmidencoding another TIE-2 ligand designated “htie-2 2” or “hTL2” isavailable under ATCC Accession No. 75928. This second ligand has beendescribed as an antagonist of the TA1-2 receptor. The identification ofsecreted human and mouse ligands for the TIE-2 receptor has beenreported by Davis et al., Cell 87, 1161-1169 (1996). The human liganddesignated “Angiopoietin-1”, to reflect its role in angiogenesis andpotential action during hemopoiesis, is the same ligand as the ligandvariously designated as “htie-2 1” or “hTL-1” in WO 96/11269.Angiopoietin-1 has been described to play an angiogenic role later anddistinct from that of VEGF (Suri et al., Cell 87, 1171-1180 (1996)).Since TIE-2 is apparently upregulated during the pathologic angiogenesisrequisite for tumor growth (Kaipainen et al., Cancer Res. 54, 6571-6577(1994)) angiopoietin-1 has been suggested to be additionally useful forspecifically targeting tumor vasculature (Davis et al., supra).

[0010] 2. PRO444

[0011] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many efforts are focused on thescreening of mammalian recombinant DNA libraries to identify the codingsequences for novel secreted proteins. We herein describe theidentification and isolation of cDNA molecules encoding novel secretedpolypeptides, designated herein as PRO444 polypeptides.

[0012] 3. PRO183

[0013] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many efforts are focused on thescreening of mammalian recombinant DNA libraries to identify the codingsequences for novel secreted proteins. We herein describe theidentification and isolation of cDNA molecules encoding novelpolypeptides, designated herein as PRO183 polypeptides.

[0014] 4. PRO185

[0015] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many efforts are focused on thescreening of mammalian recombinant DNA libraries to identify the codingsequences for novel secreted proteins. We herein describe theidentification and isolation of cDNA molecules encoding novelpolypeptides, designated herein as PRO185 polypeptides.

[0016] 5. PRO210 and PRO217

[0017] Epidermal growth factor (EGF) is a conventional mitogenic factorthat stimulates the proliferation of various types of cells includingepithelial cells and fibroblasts. EGF binds to and activates the EGFreceptor (EGFR), which initiates intracellular signaling and subsequenteffects. The EGFR is expressed in neurons of the cerebral cortex,cerebellum, and hippocampus in addition to other regions of the centralnervous system (CNS). In addition, EGF is also expressed in variousregions of the CNS. Therefore, EGF acts not only on mitotic cells, butalso on postmitotic neurons. In fact, many studies have indicated thatEGF has neurotrophic or neuromodulatory effects on various types ofneurons in the CNS. For example, EGF acts directly on cultured cerebralcortical and cerebellar neurons, enhancing neurite outgrowth andsurvival. On the other hand, EGF also acts on other cell types,including septal cholinergic and mesencephalic dopaminergic neurons,indirectly through glial cells. Evidence of the effects of EGF onneurons in the CNS is accumulating, but the mechanisms of action remainessentially unknown. EGF-induced signaling in mitotic cells is betterunderstood than in postmitotic neurons. Studies of clonedpheochromocytoma PC12 cells and cultured cerebral cortical neurons havesuggested that the EGF-induced neurotrophic actions are mediated bysustained activation of the EGFR and mitogen-activated protein kinase(MAPK) in response to EGF. The sustained intracellular signalingcorrelates with the decreased rate of EGFR down-regulation, which mightdetermine the response of neuronal cells to EGF. It is likely that EGFis a multi-potent growth factor that acts upon various types of cellsincluding mitotic cells and postmitotic neurons.

[0018] EGF is produced by the salivary and Brunner's glands of thegastrointestinal system, kidney, pancreas, thyroid gland, pituitarygland, and the nervous system, and is found in body fluids such assaliva, blood, cerebrospinal fluid (CSF), urine, amniotie fluid,prostatie fluid, pancreatie juice, and breast milk, Plata-Salaman, CRPeptides 12: 653-663 (1991).

[0019] EGF is mediated by its membrane specific receptor, which containsan intrinsic tyrosine kinase. Stoscheck CM et al., J. Cell Biochem. 31:135-152 (1986). EGF is believed to function by binding to theextracellular portion of its receptor which induces a transmembranesignal that activates the intrinsic tyrosine kinase.

[0020] Purification and sequence analysis of the EGF-like domain hasrevealed the presence of six conserved cysteine residues whichcross-bind to create three peptide loops, Savage CR et al., J. Biol.Chem. 248: 7669-7672 (1979). It is now generally known that severalother peptides can react with the EGF receptor which share the samegeneralized motif XnCX7CX4/5CX10CXCX5GX2CXn, where X represents anynon-cysteine amino acid, and n is a variable repeat number. Non isolatedpeptides having this motif include TGF-a, amphiregulin,schwannoma-derived growth factor (SDGF), heparin-binding EGF-like growthfactors and certain virally encoded peptides (e.g., Vaccinia virus,Reisner AH, Nature 313: 801-803 (1985), Shope fibroma virus, Chang W.,et al., Mol Cell Biol. 7: 535-540 (1987), Molluscum contagiosum, PorterC D & Archard L C, J. Gen. Virol. 68: 673-682 (1987), and Myxoma virus,Upton C et al., J. Virol. 61: 1271-1275 (1987). Prigent S A & Lemoine N.R., Prog. Growth Factor Res. 4: 1-24 (1992).

[0021] EGF-like domains are not confined to growth factors but have beenobserved in a variety of cell-surface and extracellular proteins whichhave interesting properties in cell adhesion, protein-proteininteraction and development, Laurence D J R & Gusterson B A, Tumor Biol.11: 229-261 (1990). These proteins include blood coagulation factors(factors VI, IX, X, XII, protein C, protein S, protein Z, tissueplasminogen activator, urokinase), extracellular matrix components(laminin, cytotactin, entactin), cell surface receptors (LDL receptor,thrombomodulin receptor) and immunity-related proteins (complement C1r,uromodulin).

[0022] Even more interesting, the general structure pattern of EGF-likeprecursors is preserved through lower organisms as well as in mammaliancells. A number of genes with developmental significance have beenidentified in invertebrates with EGF-like repeats. For example, thenotch gene of Drosophila encodes 36 tandemly arranged 40 amino acidrepeats which show homology to EGF, Wharton W et al., Cell 43: 557-581(1985). Hydropathy plots indicate a putative membrane spanning domain,with the EGF-related sequences being located on the extracellular sideof the membrane. Other homeotic genes with EGF-like repeats includeDelta, 95F and 5ZD which were identified using probes based on Notch,and the nematode gene Lin-12 which encodes a putative receptor for adevelopmental signal transmitted between two specified cells.

[0023] Specifically, EGF has been shown to have potential in thepreservation and maintenance of gastrointestinal mucosa and the repairof acute and chronic mucosal lesions, Konturek, P. C. et al., Eur. J.Gastroenterol Hepatol. 7 (10), 933-37 (1995), including the treatment ofnecrotizing enterocolitis, Zollinger-Ellison syndrome, gastrointestinalulceration gastrointestinal ulcerations and congenital microvillusatrophy, A. Guglietta & P. B. Sullivan, Eur. J. Gastroenterol Hepatol,7(10), 945-50 (1995). Additionally, EGF has been implicated in hairfollicle differentiation; C. L. du Cros, J. Invest. Dermatol. 101 (1Suppl.), 106S-113S (1993), S. G. Hillier, Clin. Endocrinol. 33(4),427-28 (1990); kidney function, L. L. Hamm et al., Semin. Nephrol. 13(1): 109-15 (1993), R. C. Harris, Am. J. Kidney Dis. 17(6): 627-30(1991); tear fluid, GB van Setten et al., Int. Ophthalmol 15(6); 359-62(1991); vitamin K mediated blood coagulation, J. Stenflo et al., Blood78(7): 1637-51 (1991). EGF is also implicated various skin diseasecharacterized by abnormal keratinocyte differentiation, e.g., psoriasis,epithelial cancers such as squamous cell carcinomas of the lung,epidermoid carcinoma of the vulva and gliomas. King, L. E. et al., Am.J. Med. Sci. 296: 154-158 (1988).

[0024] Of great interest is mounting evidence that genetie alterationsin growth factors signaling pathways are closely linked to developmentalabnormalities and to chronic diseases including cancer. Aaronson SA,Science 254: 1146-1153 (1991). For example, c-erb-2 (also known asHER-2), a proto-oncogene with close structural similarity to EGFreceptor protein, is overexpressed in human breast cancer. King et al.,Science 229: 974-976 (1985); Gullick, W. J., Hormones and their actions,Cooke BA et al., eds, Amsterdam, Elsevier, pp 349-360 (1986).

[0025] 6. PRO215

[0026] Protein-protein interactions include receptor and antigencomplexes and signaling mechanisms. As more is known about thestructural and functional mechanisms underlying protein-proteininteractions, protein-protein interactions can be more easilymanipulated to regulate the particular result of the protein-proteininteraction. Thus, the underlying mechanisms of protein-proteininteractions are of interest to the scientific and medical community.

[0027] All proteins containing leucine-rich repeats are thought to beinvolved in protein-protein interactions. Leucine-rich repeats are shortsequence motifs present in a number of proteins with diverse functionsand cellular locations. The crystal structure of ribonuclease inhibitorprotein has revealed that leucine-rich repeats correspond to beta-alphastructural units. These units are arranged so that they form a parallelbeta-sheet with one surface exposed to solvent, so that the proteinacquires an unusual, nonglubular shape. These two features have beenindicated as responsible for the protein-binding functions of proteinscontaining leucine-rich repeats. See, Kobe and Deisenhofer, TrendsBiochem. Sci., 19(10):415-421 (Oct. 1994).

[0028] A study has been reported on leucine-rich proteoglycans whichserve as tissue organizers, orienting and ordering collagen fibrilsduring ontogeny and are involved in pathological processes such as woundhealing, tissue repair, and tumor stroma formation. Iozzo, R. V., Crit.Rev. Biochem. Mol. Biol., 32(2):141-174 (1997). Others studiesimplicating leucine rich proteins in wound healing and tissue repair areDe La Salle, C., et al., Vouv. Rev. Fr. Hematol. (Germany),37(4):215-222 (1995), reporting mutations in the leucine rich motif in acomplex associated with the bleeding disorder Bernard-Soulier syndromeand Chlemetson, K. J., Thromb. Haemost. (Germany), 74(1):111-116 (July1995), reporting that platelets have leucine rich repeats. Anotherprotein of particular interest which has been reported to haveleucine-rich repeats is the SLIT protein which has been reported to beuseful in treating neuro-degenerative diseases such as Alzheimer'sdisease, nerve damage such as in Parkinson's disease, and for diagnosisof cancer, see, Artavanistsakonas, S. and Rothberg, J. M., WO9210518-A1by Yale University. Other studies reporting on the biological functionsof proteins having leucine-rich repeats include: Tayar, N., et al., Mol.Cell Endocrinol., (Ireland), 125(1-2):65-70 (December 1996)(gonadotropin receptor involvement); Miura, Y., et al., Nippon Rinsho(Japan), 54(7):1784-1789 (July 1996) (apoptosis involvement); Harris, P.C., et al., J. Am. Soc. Nephrol., 6(4):1125-1133 (October 1995) (kidneydisease involvement); and Ruoslahti, E. I., et al., WO9110727-A by LaJolla Cancer Research Foundation (decorin binding to transforming growthfactorβ involvement for treatment for cancer, wound healing andscarring).

[0029] 7. PRO242. PRO1318 and PRO1600

[0030] Leukocytes include monocytes, macrophages, basophils, andeosinophils and play an important role in the immune response. Thesecells are important in the mechanisms initiated by T and/or Blymphocytes and secrete a range of cytokines which recruit and activateother inflammatory cells and contribute to tissue destruction.

[0031] Thus, investigation of the regulatory processes by whichleukocytes move to their appropriate destination and interact with othercells is critical. Currently, leukocytes are thought to move from theblood to injured or inflamed tissues by rolling along the endothelialcells of the blood vessel wall. This movement is mediated by transientinteractions between selectins and their ligands. Next, the leukocytemust move through the vessel wall and into the tissues. This diapedesisand extravasation step involves cell activation which promotes a morestable leukocyte-endothelial cell interaction, again mediated byintegrins and their ligands.

[0032] Chemokines are a large family of structurally related polypeptidecytokines. These molecules stimulate leukocyte movement and may explainleukocyte trafficking in different inflammatory situations. Chemokinesmediate the expression of particular adhesion molecules on endothelialcells, and they produce chemoattractants which activate specific celltypes. In addition, the chemokines stimulate proliferation and regulateactivation of specific cell types. In both of these activities,chemokines demonstrate a high degree of target cell specificity.

[0033] The chemokine family is divided into two subfamilies based onwhether two amino terminal cysteine residues are immediately adjacent(C-C) or separated by one amino acid (C-X-C). Chemokines of the C-X-Cfamily generally activate neutrophils and fibroblasts while the C-Cchemokines act on a more diverse group of target cells includingmonocytes/macrophages, basophils, eosinophils and T lymphocytes. Theknown chemokines of both subfamilies are synthesized by many diversecell types as reviewed in Thomson A. (1994) The Cytokine Handbook, 2 dEd. Academic Press, N.Y.

[0034] Known chemokines include macrophage inflammatory proteins alphaand beta (MIP-1 alpha and beta ), 1-309, RANTES, and monocytechemotactie protein (MCP-1).

[0035] MIP-1 alpha and MIP-1 beta were first purified from a stimulatedmouse macrophage cell line and elicited an inflammatory response wheninjected into normal tissues. MIP-1 alpha and MIP-1 beta consist of68-69 amino acids and share approximately 70% identity in their maturesecreted forms. Both are expressed in T cells, B cells and monocyteswhich are stimulated by mitogens, anti-CD3 and endotoxin, and bothpolypeptides bind heparin and stimulate monocytes. MIP-1 alpha acts as achemoattractant for the CD-8 subset of T lymphocytes and eosinophils,while MIP-1 beta chemoattracts the CD-4 subset of T lymphocytes. Inaddition, these proteins are known to stimulate myelopoiesis in mice.

[0036] RANTES is regulated by interleukins-1 and -4, transforming nervefactor and interferon—gamma and is expressed in T cells, platelets,stimulated rheumatoid synovial fibroblasts, and in some tumor celllines. RANTES affects lymphocytes, monocytes, basophils and cosinophils.RANTES expression is substantially reduced upon T cell stimulation.

[0037] Monocyte chemotactie protein (MCP-1) is a 76 amino acid proteinwhich appears to be expressed in almost all cells and tissues uponstimulation by a variety of agents. However, the targets of MCP-1 arelimited to monocytes and basophils. In these cells, MCP-1 induces aMCP-1 receptor. Two related proteins, MCP-2 and MCP-3, have 62% and 73%identity, respectively, with MCP-1 and share its chemoattractantspecificity or monocytes.

[0038] Current techniques for diagnosis of abnormalities in inflamed ordiseased issues mainly rely on observation of clinical symptoms orserological analyses of body tissues or fluids for hormones,polypeptides or various metabolites. Problems exist with thesediagnostic techniques. First, patients may not manifest clinicalsymptoms at early stages of disease. Second, serological tests do notalways differentiate between invasive diseases and genetie syndromes.Thus, the identification of expressed chemokines is important to thedevelopment of new diagnostic techniques, effective therapies, and toaid in the understanding of molecular pathogenesis.

[0039] The chemokine molecules were reviewed in Schall TJ (1994)Chemotactie Cytokines: Targets for Therapeutie Development.International Business Communications, Southborough Mass. pp 180-270;and in Paul WE (1993) Fundamental Immunology, 3rd Ed. Raven Press, N.Y.pp 822-826.

[0040]8. PRO288

[0041] Control of cell numbers in mammals is believed to be determined,in part, by a balance between cell proliferation and cell death. Oneform of cell death, sometimes referred to as necrotic cell death, istypically characterized as a pathologic form of cell death resultingfrom some trauma or cellular injury. In contrast, there is another,“physiologic” form of cell death which usually proceeds in an orderly orcontrolled manner. This orderly or controlled form of cell death isoften referred to as “apoptosis” [see, e.g., Barr et al.,Bio/Technology, 12:487-493 (1994); Steller et al., Science,267:1445-1449 (1995)]. Apoptotie cell death naturally occurs in manyphysiological processes, including embryonic development and clonalselection in the immune system [Itoh et al., Cell, 66:233-243 (1991)].Decreased levels of apoptotie cell death have been associated with avariety of pathological conditions, including cancer, lupus, and herpesvirus infection [Thompson, Science, 267:1456-1462 (1995)]. Increasedlevels of apoptotie cell death may be associated with a variety of otherpathological conditions, including AIDS, Alzheimer's disease,Parkinson's disease, amyotrophic lateral sclerosis, multiple sclerosis,retinitis pigmentosa, cerebellar degeneration, aplastic anemia,myocardial infarction, stroke, reperfusion injury, and toxin-inducedliver disease [see, Thompson, supra].

[0042] Apoptotie cell death is typically accompanied by one or morecharacteristic morphological and biochemical changes in cells, such ascondensation of cytoplasm, loss of plasma membrane microvilli,segmentation of the nucleus, degradation of chromosomal DNA or loss ofmitochondrial function. A variety of extrinsic and intrinsic signals arebelieved to trigger or induce such morphological and biochemicalcellular changes [Raff, Nature, 356:397-400 (1992); Steller, supra;Sachs et al., Blood, 82:15 (1993)]. For instance, they can be triggeredby hormonal stimuli, such as glucocorticoid hormones for immaturethymocytes, as well as withdrawal of certain growth factors[Watanabe-Fukunaga et al., Nature, 356:314-317 (1992)]. Also, someidentified oncogenes such as myc, rel, and E1A, and tumor suppressors,like p53, have been reported to have a role in inducing apoptosis.Certain chemotherapy drugs and some forms of radiation have likewisebeen observed to have apoptosis-inducing activity [Thompson, supra].

[0043] Various molecules, such as tumor necrosis factor-α(“TNF-α”),tumor necrosis factor-β (“TNF-β” or “lymphotoxin”), CD30 ligand, CD27ligand, CD40 ligand, OX-40 ligand, 4-1BB ligand, Apo-1 ligand (alsoreferred to as Fas ligand or CD95 ligand), and Apo-2 ligand (alsoreferred to as TRAIL) have been identified as members of the tumornecrosis factor (“TNF”) family of cytokines [See, e.g., Gruss and Dower,Blood, 85:3378-3404 (1995); Wiley et al., Immunity, 3:673-682 (1995);Pitti et al., J. Biol. Chem., 271:12687-12690 (1996)]. Among thesemolecules, TNF-α, TNF-β, CD30 ligand, 4-1BB ligand, Apo-1 ligand, andApo-2 ligand (TRAIL) have been reported to be involved in apoptotie celldeath. Both TNF-α and TNF-β have been reported to induce apoptotie deathin susceptible tumor cells [Schmid et al., Proc. Natl. Acad. Sci.,83:1881 (1986); Dealtry et al., Eur. J. Immunol., 17:689 (1987)]. Zhenget al. have reported that TNF-α is involved in post-stimulationapoptosis of CD8-positive T cells [Zheng et al., Nature, 377:348-351(1995)]. Other investigators have reported that CD30 ligand may beinvolved in deletion of self-reactive T cells in the thymus [Amakawa etal., Cold Spring Harbor Laboratory Symposium on Programmed Cell Death,Abstr. No. 10, (1995)].

[0044] Mutations in the mouse Fas/Apo-1 receptor or ligand genes (calledlpr and gld, respectively) have been associated with some autoimmunedisorders, indicating that Apo-1 ligand may play a role in regulatingthe clonal deletion of self-reactive lymphocytes in the periphery[Krammer et al., Curr. Op. Immunol., 6:279-289 (1994); Nagata et al.,Science, 267:1449-1456 (1995)]. Apo-1 ligand is also reported to inducepost-stimulation apoptosis in CD4-positive T lymphocytes and in Blymphocytes, and may be involved in the elimination of activatedlymphocytes when their function is no longer needed [Krammer et al.,supra; Nagata et al., supra]. Agonist mouse monoclonal antibodiesspecifically binding to the Apo-1 receptor have been reported to exhibitcell killing activity that is comparable to or similar to that of TNF-α[Yonehara et al., J. Exp. Med., 169:1747-1756 (1989)].

[0045] Induction of various cellular responses mediated by such TNFfamily cytokines is believed to be initiated by their binding tospecific cell receptors. Two distinct TNF receptors of approximately55-kDa (TNFR 1) and 75-kDa (TNFR2) have been identified [Hohman et al.,J. Biol. Chem., 264:14927-14934 (1989); Brockhaus et al., Proc. Natl.Acad. Sci., 87:3127-3131 (1990); EP 417,563, published Mar. 20, 1991]and human and mouse cDNAs corresponding to both receptor types have beenisolated and characterized [Loetscher et al., Cell, 61:351 (1990);Schall et al., Cell, 61:361 (1990); Smith et al., Science, 248:1019-1023(1990); Lewis et al., Proc. Natl. Acad. Sci., 88:2830-2834 (1991);Goodwin et al., Mol. Cell. Biol., 11:3020-3026 (1991)]. Extensivepolymorphisms have been associated with both TNF receptor genes [see,e.g., Takao et al., Immunogenetics, 37:199-203 (1993)]. Both TNFRs sharethe typical structure of cell surface receptors including extracellular,transmembrane and intracellular regions. The extracellular portions ofboth receptors are found naturally also as soluble TNF-binding proteins[Nophar, Y. et al., EMBO J., 2:3269 (1990); and Kohno, T. et al., Proc.Natl. Acad. Sci. U.S.A., 87:8331 (1990)]. More recently, the cloning ofrecombinant soluble TNF receptors was reported by Hale et al. [J. Cell.Biochem. Supplement 15F, 1991, p. 113 (P424)].

[0046] The extracellular portion of type 1 and type 2 TNFRs (TNFR1 andTNFR2) contains a repetitive amino acid sequence pattern of fourcysteine-rich domains (CRDs) designated 1 through 4, starting from theNH₂-terminus. Each CRD is about 40 amino acids long and contains 4 to 6cysteine residues at positions which are well conserved [Schall et al.,supra; Loctscher et al., supra; Smith et al., supra; Nophar et al.,supra; Kohno et al., supra]. In TNFR1, the approximate boundaries of thefour CRDs areas follows: CRD1-amino acids 14 to about 53; CRD2-aminoacids from about 54 to about 97; CRD3-amino acids from about 98 to about138; CRD4-amino acids from about 139 to about 167. In TNFR2, CRD1includes amino acids 17 to about 54; CRD2-amino acids from about 55 toabout 97; CRD3-amino acids from about 98 to about 140; and CRD4-aminoacids from about 141 to about 179 [Banner et al., Cell, 73:431-435(1993)]. The potential role of the CRDs in ligand binding is alsodescribed by Banner et al., supra.

[0047] A similar repetitive pattern of CRDs exists in several othercell-surface proteins, including the p75 nerve growth factor receptor(NGFR) [Johnson et al., Cell, 47:545 (1986); Radeke et al., Nature,325:593 (1987)], the B cell antigen CD40 [Stamenkovic et al., EMBO J.,8:1403 (1989)], the T cell antigen OX40 [Mallet et al., EMBO J., 9:1063(1990)] and the Fas antigen [Yonehara et al., supra and Itoh et al.,supra]. CRDs are also found in the soluble TNFR (sTNFR)-like T2 proteinsof the Shope and myxoma poxviruses [Upton et al., Virology, 160:20-29(1987); Smith et al., Biochem. Biophys. Res. Commun., 176:335 (1991);Upton et al., Virology, 184:370 (1991)]. Optimal alignment of thesesequences indicates that the positions of the cysteine residues are wellconserved. These receptors are sometimes collectively referred to asmembers of the TNF/NGF receptor superfamily. Recent studies on p75NGFRshowed that the deletion of CRD1 [Welcher, A.A. et al., Proc. Natl.Acad. Sci. USA, 88:159-163 (1991)] or a 5-amino acid insertion in thisdomain [Yan, H. and Chao, M. V., J. Biol. Chem., 266:12099-12104 (1991)]had little or no effect on NGF binding [Yan, H. and Chao, M. V., supra].p75 NGFR contains a proline-rich stretch of about 60 amino acids,between its CRD4 and transmembrane region, which is not involved in NGFbinding [Peetre, C. et al., Eur. J. Hematol., 41:414-419 (1988);Seckinger, P. et al., J. Biol. Chem., 264:11966-11973 (1989); Yan, H.and Chao, M. V., supra]. A similar proline-rich region is found in TNFR2but not in TNFR1.

[0048] Itoh et al. disclose that the Apo-1 receptor can signal anapoptotie cell death similar to that signaled by the 55-kDa TNFR1 [Itohet al., supra]. Expression of the Apo-1 antigen has also been reportedto be down-regulated along with that of TNFR1 when cells are treatedwith either TNF-α or anti-Apo-1 mouse monoclonal antibody [Krammer etal., supra; Nagata et al., supra]. Accordingly, some investigators havehypothesized that cell lines that co-express both Apo-1 and TNFR1receptors may mediate cell killing through common signaling pathways[Id.].

[0049] The TNF family ligands identified to date, with the exception oflymphotoxin-α, are type II transmembrane proteins, whose C-terminus isextracellular. In contrast, the receptors in the TNF receptor (TNFR)family identified to date are type I transmembrane proteins. In both theTNF ligand and receptor families, however, homology identified betweenfamily members has been found mainly in the extracellular domain(“ECD”). Several of the TNF family cytokines, including TNF-α, Apo-1ligand and CD40 ligand, are cleaved proteolytically at the cell surface;the resulting protein in each case typically forms a homotrimericmolecule that functions as a soluble cytokine. TNF receptor familyproteins are also usually cleaved proteolytically to release solublereceptor ECDs that can function as inhibitors of the cognate cytokines.

[0050] Recently, other members of the TNFR family have been identified.In Marsters et al., Curr. Biol., 6:750 (1996), investigators describe afull length native sequence human polypeptide, called Apo-3, whichexhibits similarity to the TNFR family in its extracellularcysteine-richi repeats and resembles TNFR1 and CD95 in that it containsa cytoplasinic death domain sequence [see also Marsters et al., Curr.Biol., 6:1669 (1996)]. Apo-3 has also been referred to by otherinvestigators as DR3, ws1-1 and TRAMP [Chinnaiyan et al., Science,274:990 (1996); Kitson et al., Nature, 384:372 (1996); Bodmer et al.,Immunity, 6:79 (1997)].

[0051] Pan et al. have disclosed another TNF receptor family memberreferred to as “DR4” [Pan et al., Science, 276:111-113 (1997)]. The DR4was reported to contain a cytoplasmic death domain capable of engagingthe cell suicide apparatus. Pan et al. disclose that DR4 is believed tobe a receptor for the ligand known as Apo-2 ligand or TRAIL.

[0052] In Sheridan et al., Science, 277:818-821 (1997) and Pan et al.,Science, 277:815-818 (1997), another molecule believed to be a receptorfor the Apo-2 ligand (TRAIL) is described. That molecule is referred toas DR5 (it has also been alternatively referred to as Apo-2). Like DR4,DR5 is reported to contain a cytoplasmic death domain and be capable ofsignaling apoptosis.

[0053] In Sheridan et al., supra, a receptor called DcR1 (oralternatively, Apo-2DcR) is disclosed as being a potential decoyreceptor for Apo-2 ligand (TRAIL). Sheridan et al. report that DcR1 caninhibit Apo-2 ligand function in vitro. See also, Pan et al., supra, fordisclosure on the decoy receptor referred to as TRID.

[0054] As presently understood, the cell death program contains at leastthree important elements—activators, inhibitors, and effectors; in C.elegans, these elements are encoded respectively by three genes, Ced-4,Ced-9 and Ced-3 [Steller, Science, 267:1445 (1995); Chinnaiyan et al.,Science, 275:1122-1126 (1997); Wang et al., Cell, 90:1-20 (1997)]. Twoof the TNFR family members, TNFR1 and Fas/Apo1 (CD95), can activateapoptotie cell death [Chinnaiyan and Dixit, Current Biolopy, 6:555-562(1996); Fraser and Evan, Cell; 85:781-784 (1996)]. TNFR1 is also knownto mediate activation of the transcription factor, NF-κB [Tartaglia etal., Cell, 74:845-853 (1993); Hsu et al., Cell, 84:299-308 (1996)]. Inaddition to some ECD homology, these two receptors share homology intheir intracellular domain (ICD) in an oligomerization interface knownas the death domain [Tartaglia et al., supra; Nagata, Cell, 88:355(1997)]. Death domains are also found in several metazoan proteins thatregulate apoptosis, namely, the Drosophila protein, Reaper, and themammalian proteins referred to as FADD/MORT1, TRADD, and RIP [Cleavelandand Ihle, Cell, 81:479-482 (1995)]. Using the yeast-two hybrid system,Raven et al. report the identification of protein, wsl-1, which binds tothe TNFR1 death domain [Raven et al., Programmed Cell Death Meeting,Sep. 20-24, 1995, Abstract at page 127; Raven et al., European CytokineNetwork, 7:Abstr. 82 at page 210 (April-June 1996); see also, Kitson etal., Nature, 384:372-375 (1996)]. The wsl-1 protein is described asbeing homologous to TNFR1 (48% identity) and having a restricted tissuedistribution. According to Raven et al., the tissue distribution ofwsl-1 is significantly different from the TNFR1 binding protein, TRADD.

[0055] Upon ligand binding and receptor clustering, TNFR1 and CD95 arebelieved to recruit FADD into a death-inducing signalling complex. CD95purportedly binds FADD directly, while TNFR1 binds FADD indirectly viaTRADD [Chinnaiyan et al., Cell, 81:505-512 (1995); Boldin et al., J.Biol. Chem., 270:387-391 (1995); Hsu et al., supra; Chinnaiyan et al.,J. Biol. Chem., 271:4961-4965 (1996)]. It has been reported that FADDserves as an adaptor protein which recruits the Ced-3-related protease,MACHα/FLICE (caspase 8), into the death signalling complex [Boldin etal., Cell, 85:803-815 (1996); Muzio et al., Cell, 85:817-827 (1996)].MACHα/FLICE appears to be the trigger that sets off a cascade ofapoptotie proteases, including the interleukin-1β converting enzyme(ICE) and CPP32/Yama, which may execute some critical aspects of thecell death programme [Fraser and Evan, supra].

[0056] It was recently disclosed that programmed cell death involves theactivity of members of a family of cysteine proteases related to the C.elegans cell death gene, ced-3, and to the mammalian IL-1-convertingenzyme, ICE. The activity of the ICE and CPP32/Yama proteases can beinhibited by the product of the cowpox virus gene, crnA [Ray et al.,Cell, 69:597-604 (1992); Tewari et al., Cell, 81:801-809 (1995)]. Recentstudies show that CrmA can inhibit TNFR1-and CD95-induced cell death[Enari et al., Nature, 375:78-81 (1995); Tewari et al., J. Biol. Chem.,270:3255-3260 (1995)].

[0057] As reviewed recently by Tewari et al., TNFR1, TNFR2 and CD40modulate the expression of proinflammatory and costimulatory cytokines,cytokine receptors, and cell adhesion molecules through activation ofthe transcription factor, NF-κB [Tewari et al., Curr. Op. Genet.Develop., 6:39-44 (1996)]. NF-κB is the prototype of a family of dimerictranscription factors whose subunits contain conserved Rel regions[Verma et al., Genes Develop., 9:2723-2735 (1996); Baldwin, Ann. Rev.Immunol., 14:649-681 (1996)]. In its latent form, NF-κB is complexedwith members of the IκB inhibitor family; upon inactivation of the IκBin response to certain stimuli, released NF-κB translocates to thenucleus where it binds to specific DNA sequences and activates genetranscription.

[0058] For a review of the TNF family of cytokines and their receptors,see Gruss and Dower, supra.

[0059] 9. PRO365

[0060] Polypeptides such as human 2-19 protein may function ascytokines. Cytokines are low molecular weight proteins which function tostimulate or inhibit the differentiation, proliferation or function ofimmune cells. Cytokines often act as intercellular messengers and havemultiple physiological effects. Given the physiological importance ofimmune mechanisms in vivo, efforts are currently being under taken toidentify new, native proteins which are involved in effecting the immunesystem. We describe herein the identification of a novel polypeptidewhich has homology to the human 2-19 protein.

[0061] 10. PRO1361

[0062] Efforts are being undertaken by both industry and academia toidentify new, native transmembrane receptor proteins. Many efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel receptor proteins. We hereindescribe the identification and characterization of novel transmembranepolypeptides, designated herein as PRO1361 polypeptides.

[0063] 11. PRO1308

[0064] Follistatin is a secreted protein that regulates secretion ofpituitary follicle-stimulating hormone (FSH). It functions by bindingto, and thereby inhibiting, proteins such as activin and other membersof the transforming growth factor beta (TGFβ) family, that stimulate theproduction and secretion of FSH from the anterior pituitary. Follistatinis also involved in mechanisms that control basic development, includingthe induction of neural development. Follistatin also exhibitsangiogenic properties, particularly in combination with basic fibroblastgrowth factor (bFGF). As such, there is strong interest in identifyingnew members of the follistatin family of proteins. The identificationand characterization of follistatins is the topic of the followingreferences which are incorporated herein by reference: Sugino et al. J.Med Invest (1997) 44:(1-2): 1-14; Mather et al., Proc. Soc. Exp. biol.Med. (1997) 215(3):209-222; Thomsen, G. H., Trends Genet (1997) 13(6):209-211; DePaolo, L. V., Proc. Soc. Exp. Biol. Med. (1997)214(4):328-339; Peng et al., Biol. Signals (1996) 5(2):81-89, andHalvorson et al. Fertil Steril (1996) 65(3):459-469.

[0065] 12. PRO1183

[0066] Protoporphyrinogen oxidase catalyzes the penultimate step in theheme biosynthetic pathway. Deficiency in activity of this enzyme resultsin the human genetie disease variegate porphyria. Thus,protoporphyrinogen oxidases and molecules which either modulate or arerelated to these oxidases are of interest. Moreover, oxidases, andrelated molecules in general are also of interest. Oxidases are furtherdescribed in at least Birchfield, et al., Biochemistry, 37(19):6905-6910(1998); Fingar, et al., Cancer Res., 57(20):4551-4556 (1997); Arnould,et al., Biochemistry, 36(33):10178-10184 (1997); Cell Mol. Biol.,43(1):67-73 (1997).

[0067] 13. PRO1272

[0068] The cement gland is an ectodermal organ in the head of frogembryos, lying anterior to any neural tissue. The cement gland, likeneural tissue, has been shown to be induced by the dorsal mesoderm.XAG-1 is a cement gland specific protein that is useful as a marker ofcement gland induction during development. See, Sive, et al., Cell,58(1):171-180 (1989); Itoh, et al., Development, 121(12):3979-3988(1995). XAG-2 and other proteins related to the XAG family are furtherdescribed in Aberger, et al., Mech. Dev., 72(1-2):115-130 (1998) andGammill and Sive, Development, 124(2):471-481 (1997). Thus, novelpolypeptides having sequence identity with XAG proteins are of interest.

[0069] 14. PRO1419

[0070] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many efforts are focused on thescreening of mammalian recombinant DNA libraries to identify the codingsequences for novel secreted proteins. We herein describe theidentification and characterization of novel secreted polypeptides,designated herein as PRO1419 polypeptides.

[0071] 15. PRO4999

[0072] Uromodulin is synthesized in the kidney and is the most abundantprotein in normal human urine. The amino acid sequence encoded by one ofthe exons of the uromodulin gene has homology to thelow-density-lipoprotein receptor and the epidermal growth factorprecursor. Pennica et al., Science 236:83-88 (1987). The function ofuromodulin is not known; however, it may function as a unique renalregulatory glycoprotein that specifically binds to and regulates thecirculating activity of a number of potent cytokines, as it binds toIL-1, IL-2 and TNF with high affinity. See Hession et al., Science237:1479-1484 (1987). Su et al. suggest that uromodulin plays asignificant role in the innate immunity of the urinary system and thatthe immunostimulatory activity of uromodulin is potentially useful forimmunotherapy. Su et al., J. Immunology, 158:3449-3456 (1997).

[0073] We herein describe the identification and characterization ofnovel polypeptides having sequence similarity to uromodulin, designatedherein as PRO4999 polypeptides.

[0074] 16. PRO7170

[0075] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many efforts are focused on thescreening of mammalian recombinant DNA libraries to identify the codingsequences for novel secreted proteins. We herein describe theidentification and characterization of novel secreted polypeptides,designated herein as PRO7170 polypeptides.

[0076] 17. PRO248

[0077] Cytokines have been implicated in the pathogenesis of a number ofbrain diseases in which neurological dysfuntion has been attributed to achange in amino acid neurotransmitter metabolism. In particular, membersof the transforming growth factorβs (TGFβ) have been implicated.Transforming growth peptides are small polypeptides that were firstidentified by their ability to induce proliferation and transformationin noncancerous cells in culture. Although initially defined as a growthfactor, TGFβ also inhibits proliferation of epithelial, endothelial,lymphoid, and hematopoietic cells. This cytokine is thought to play animportant role in regulating the duration of the inflammatory response,allowing the healing process to proceed. It is also a potentimmunomodulator, which has many pleiotrophic effects, includingregulating many other cytokines.

[0078] The TGFβ family includes basic myelin proteins (BMP-2, BMP-4,BMP-5, BMP-6, BMP-7), activins A & B, decapentaplegic (dpp), 60A, OP-2,dorsalin, growth differentiation factors (GDFs) 1, 3, and 9, nodal, MIS,Inhibin α, transforming growth factors betas (TGF-62 1, TGF-β2, TGF-β3,TGF-β5), and glial-derived neurotrophic factor (GDNF), Atrisano, et al.,J. Biochemica et Biophysica Acta. 1222:71-80 (1994). Of particuarinterest are the growth differentiation factors, for as their nameimplies, these factors are implicated in the differentiation of cells.

[0079] Thus, identifying proteins having homology to the TGFβ familymembers, particularly growth differentiation factor (GDF)3, is ofimportance to the medical and industrial community. Generally, proteinshaving homology to each other have similar function. It is also ofinterest when proteins having homology do not have similar functions,indicating that certain structural motifs identify information otherthan function, such as locality of function.

[0080] 18. PRO353

[0081] The complement proteins comprise a large group of serum proteinssome of which act in an enzymatic cascade, producing effector moleculesinvolved in inflammation. The complement proteins are of particularimportance in regulating movement and function of cells involved ininflammation. Given the physiological importance of inflammation andrelated mechanisms in vivo, efforts are currently being under taken toidentify new, native proteins which are involved in inflammation. Wedescribe herein the identification and characterization of novelpolypeptides which have homology to complement proteins, designatedherein as PRO353 polypeptides.

[0082] 19. PRO533

[0083] Growth factors are molecular signals or mediators that enhancecell growth or proliferation, alone or in concert, by binding tospecific cell surface receptors. however, there are other cellularreactions than only growth upon expression to growth factors. As aresult, growth factors are better characterized as multifunctional andpotent cellular regulators. Their biological effects includeproliferation, chemotaxis and stimulation of extracellular matrixproduction. Growth factors can have both stimulatory and inhibitoryeffects. For example, transforming growth factors (TGF-β) is highlypleiotropic and can stimulate proliferation in some cells, especiallyconnective tissues, while being a potent inhibitor of proliferation inothers, such as lymphocytes and epithelial cells.

[0084] The physiological effect of growth stimulation or inhibition bygrowth factors depends upon the state of development and differentiationof the target tissue. The mechanism of local cellular regulation byclassical endocrine molecules comprehends autocrine (same cell),juxtacrine (neighbor cell), and paracrine (adjacent cell) pathways.Peptide growth factors are elements of a complex biological language,providing the basis for intercellular communication. They permit cellsto convey information between each other, mediate interaction betweencells and change gene expression. the effect of these multifunctionaland pluripotent factors is dependent on the presence or absence of otherpeptides.

[0085] Fibroblast growth factors (FGFs) are a family of heparin-binding,potent mitogens for both normal diploid fibroblasts and established celllines, Godpodarowicz, D. et al. (1984), Proc. Natl. Acad. Sci. USA 81:6983. the FGF family comprises acidic FGF (FGF-1), basic FGF (FGF-2),INT-2 (FGF-3), K-FGF/HST (FGF-4), FGF-5, FGF-6, KGF (FGF-7), AIGF(FGF-8) among others. All FGFs have two conserved cysteine residues andshare 30-50% sequence homology at the amino acid level. These factorsare mitogenic for a wide variety of normal diploid mesoderm-derived andneural crest-derived cells, inducing granulosa cells, adrenal corticalcells, chrondocytes, myoblasts, corneal and vascular endothelial cells(bovine or human), vascular smooth muscle cells, lens, retina andprostatic epithelial cells, oligodendrocytes, astrocytes, chrondocytes,myoblasts and osteoblasts.

[0086] Fibroblast growth factors can also stimulate a large number ofcell types in a non-mitogenic manner. These activities include promotionof cell migration into a wound area (chemotaxis), initiation of newblood vessel formulation (angiogenesis), modulation of nerveregeneration and survival (neurotrophism), modulation of endocrinefunctions, and stimulation or suppression of specific cellular proteinexpression, extracellular matrix production and cell survival. Baird, A.& Bohlen, P., Handbook of Exp. Phrmacol. 9(1): 369-418 (1990). Theseproperties provide a basis for using fibroblast growth factors intherapeutie approaches to accelerate wound healing, nerve repair,collateral blood vessel formation, and the like. For example, fibroblastgrowth factors, have been suggested to minimize myocardium damage inheart disease and surgery (U.S. Pat. No. 4,378,437).

[0087] We herein describe the identification and characterization ofnovel polypeptides having homology to FGF, herein designated PRO533polypeptides.

[0088] 20. PRO301

[0089] The widespread occurrence of cancer has prompted the devotion ofconsiderable resources and discovering new treatments of treatment. Oneparticular method involves the creation of tumor or cancer specificmonoclonal antibodies (mAbs) which are specific to tumor antigens. SuchmAbs, which can distinguish between normal and cancerous cells areuseful in the diagnosis, prognosis and treatment of the disease.Particular antigens are known to be associated with neoplastic diseases,such as colorectal cancer.

[0090] One particular antigen, the A33 antigen is expressed in more than90% of primary or metastatic colon cancers as well as normal colonepithelium. Since colon cancer is a widespread disease, early diagnosisand treatment is an important medical goal. Diagnosis and treatment ofcolon cancer can be implemented using monoclonal antibodies (mAbs)specific therefore having fluorescent, nuclear magnetic or radioactivetags. Radioactive gene, toxins and/or drug tagged mAbs can be used fortreatment in situ with minimal patient description. mabs can also beused to diagnose during the diagnosis and treatment of colon cancers.For example, when the serum levels of the A33 antigen are elevated in apatient, a drop of the levels after surgery would indicate the tumorresection was successful. On the other hand, a subsequent rise in serumA33 antigen levels after surgery would indicate that metastases of theoriginal tumor may have formed or that new primary tumors may haveappeared. Such monoclonal antibodies can be used in lieu of, or inconjunction with surgery and/or other chemotherapies. For example, U.S.Pat. No. 4,579,827 and U.S. Ser. No. 424,991 (E.P. 199,141) are directedto therapeutie administration of monoclonal antibodies, the latter ofwhich relates to the application of anti-A33 mAb.

[0091] Many cancers of epithelial origin have adenovirus receptors. Infact, adenovirus-derived vectors have been proposed as a means ofinserting antisense nucleic acids into tumors (U.S. Pat. No. 5,518,885).Thus, the association of viral receptors with neoplastic tumors is notunexpected.

[0092] We herein describe the identification and characterization ofnovel polypeptides having homology to certain cancer-associatedantigens, designated herein as PRO301 polypeptides.

[0093] 21. PRO187

[0094] Growth factors are molecular signals or mediators that enhancecell growth or proliferation, alone or in concert, by binding tospecific cell surface receptors. However, there are other cellularreactions than only growth upon expression to growth factors. As aresult, growth factors are better characterized as multifunctional andpotent cellular regulators. Their biological effects includeproliferation, chemotaxis and stimulation of extracellular matrixproduction. Growth factors can have both stimulatory and inhibitoryeffects. For example, transforming growth factor (TGF-β) is highlypleiotropic and can stimulate proliferation in some cells, especiallyconnective tissue, while being a potent inhibitor of proliferation inothers, such as lymphocytes and epithelial cells.

[0095] The physiological effect of growth stimulation or inhibition bygrowth factors depends upon the state of development and differentiationof the target tissue. The mechanism of local cellular regulation byclassical endocrine molecules involves comprehends autocrine (samecell), juxtacrine (neighbor cell), and paracrine (adjacent cells)pathways. Peptide growth factors are elements of a complex biologicallanguage, providing the basis for intercellular communication. Theypermit cells to convey information between each other, mediateinteraction between cells and change gene expression. The effect ofthese multifunctional and pluripotent factors is dependent on thepresence or absence of other peptides.

[0096] FGF-8 is a member of the fibroblast growth factors (FGFs) whichare a family of heparin-binding, potent mitogens for both normal diploidfibroblasts and established cell lines, Gospodarowicz et al. (1984),Proc. Natl. Acad. Sci. USA 81:6963. The FGF family comprises acidic FGF(FGF-1), basic FGF (FGF-2), INT-2 (FGF-3), K-FGF/HST (FGF-4), FGF-5,FGF-6, KGF (FGF-7), AIGF (FGF-8) among others. All FGFs have twoconserved cysteine residues and share 30-50% sequence homology at theamino acid level. These factors are mitogenic for a wide variety ofnormal diploid mesoderm-derived and neural crest-derived cells,including granulosa cells, adrenal cortical cells, chondrocytes,inyoblasts, corneal and vascular endothelial cells (bovine or human),vascular smooth muscle cells, lens, retina and prostatic epithelialcells, oligodendrocytes, astrocytes, chrondocytes, myoblasts andosteoblasts.

[0097] Fibroblast growth factors can also stimulate a large number ofcell types in a non-mitogenic manner. These activities include promotionof cell migration into wound area (chemotaxis), initiation of new bloodvessel formulation (angiogenesis), modulation of nerve regeneration andsurvival (neurotrophism), modulation of endocrine functions, andstimulation or suppression of specific cellular protein expression,extracellular matrix production and cell survival. Baird & Bohlen,Handbook of Exp. Pharmacol. 95(1): 369-418, Springer, (1990). Theseproperties provide a basis for using fibroblast growth factors intherapeutie approaches to accelerate wound healing, nerve repair,collateral blood vessel formation, and the like. For example, fibroblastgrowth factors have been suggested to minimize myocardium damage inheart disease and surgery (U.S. Pat. No. 4,378,347).

[0098] FGF-8, also known as androgen-induced growth factor (AIGF), is a215 amino acid protein which shares 30-40% sequence homology with theother members of the FGF family. FGF-8 has been proposed to be underandrogenic regulation and induction in the mouse mammary carcinoma cellline SC3. Tanaka et al., Proc. Natl. Acad. Sci. USA 89: 8928-8932(1992); Sato et al., J. Steroid Biochem. Molec. Biol. 47: 91-98 (1993).As a result, FGF-8 may have a local role in the prostate, which is knownto be an androgen-responsive organ. FGF-8 can also be oncogenic, as itdisplays transforming activity when transfected into NIH-3T3fibroblasts. Kouhara et al., Oncogene 9 455-462 (1994). While FGF-8 hasbeen detected in heart, brain, lung, kidney, testis, prostate and ovary,expression was also detected in the absence of exogenous androgens.Schmitt et al., J. Steroid Biochein. Mol. Biol. 57 (3-4): 173-78 (1996).

[0099] FGF-8 shares the property with several other FGFs of beingexpressed at a variety of stages of murine embryogenesis, which supportsthe theory that the various FGFs have multiple and perhaps coordinatedroles in differentiation and embryogenesis. Moreover, FGF-8 has alsobeen identified as a protooncogene that cooperates with Wnt-1 in theprocess of mammary tumorigenesis (Shackleford et al., Proc. Natl. Acad.Sci. USA 90, 740-744 (1993); Heikinheimo et al., Mech. Dev. 48: 129-138(1994)).

[0100] In contrast to the other FGFs, FGF-8 exists as three proteinisoforms, as a result of alternative splicing of the primary transcript.Tanaka et al., supra. Normal adult expression of FGF-8 is weak andconfined to gonadal tissue, however northern blot analysis has indicatedthat FGF-8 mRNA is present from day 10 through day 12 or murinegestation, which suggests that FGF-8 is important to normal development.Heikinheimo et al., Mech Dev. 48(2):129-38 (1994). Further in situhybridization assays between day 8 and 16 of gestation indicated initialexpression in the surface ectoderm of the first bronchial arches, thefrontonasal process, the forebrain and the midbrain-hindbrain junction.At days 10-12, FGF-8 was expressed in the surface ectoderm of theforelimb and hindlimb buds, the nasal its and nasopharynx, theinfundibulum and in the telencephalon, diencephalon and metencephalon.Expression continues in the developing hindlimbs through day 13 ofgestation, but is undetectable thereafter. The results suggest thatFGF-8 has a unique temporal and spatial pattern in embryogenesis andsuggests a role for this growth factor in multiple regions of ectodermaldifferentiation in the post-gastrulation embryo.

[0101] We herein describe the identification of novel poypeptides havinghomology to FGF-8, wherein those polypeptides are heein designatedPRO187 polypeptides.

[0102] 22. PRO337

[0103] Neuronal development in higher vertebrates is characterized byprocesses that must successfully navigate distinct cellular environmenten route to their synaptic targets. The result is a functionally preciseformation of neural circuits. The precision is believed to result formmechanisms that regulate growth cone pathfinding and target recognition,followed by latter refinement and remodeling of such projections byevents that require neuronal activity, Goodman and Shatz, Cell/Neuron[Suppl.] 72(10): 77-98 (1993). It is further evident that differentneurons extend nerve fibers that are biochemically distinct and rely onspecific guidance cues provided by cell-cell, cell-matrix, andchemotrophic interactions to reach their appropriate synaptic targets,Goodman et al., supra.

[0104] One particular means by which diversity of the neuronal cellsurface may be generated is through differential expression of cellsurface proteins referred to as cell adhesion molecules (CAMs).Neuronally expressed CAMs have been implicated in diverse developmentalprocesses, including migration of neurons along radial glial cells,providing permissive or repulsive substrates for neurite extension, andin promoting the selective fasciculation of axons in projectionalpathways. Jessel, Neuron 1: 3-13 (1988); Edelman and Crossin, Annu. Rev.Biochem. 60: 155-190 (1991). Interactions between CAMs present on thegrowth cone membrane and molecules on opposing cell membranes or in theextracellular matrix are thought to provide the specific guidance cuesthat direct nerve fiber outgrowth along appropriate projectionalpathways. Such interactions are likely to result in the activation ofvarious second messenger systems within the growth cone that regulateneurite outgrowth. Doherty and Walsh, Curr. Opin Neurobiol. 2: 595-601(1992).

[0105] In higher vertebrates, most neural CAMs have been found to bemembers of three major structural families of proteins: the integrins,the cadherins, and the immunoglobulin gene superfamily (IgSF). Jessel,supra.; Takeichi, Annu. Rev. Biochein. 59: 237-252 (1990); Reichardt andTomaselli, Annu. Rev. Neurosci. 14: 531-570 (1991). Cell adhesionmolecules of the IgSF (or Ig-CAMs), in particular, constitute a largefamily of proteins frequently implicated in neural cell interactions andnerve fiber outgrowth during development, Salzer and Colman, Dev.Neurosci. 11: 377-390 (1989); Brummendorf and Rathjen, J. Neurochem. 61:1207-1219 (1993). However, the majority of mammalian Ig-CAMs appear tobe too widely expressed to specify navigational pathways or synaptictargets suggesting that other CAMs, yet to be identified, have role inthese more selective interactions of neurons.

[0106] Many of the known neural Ig-CAMs have been found to be attachedto the plasma membrane via a glycosylphosphatidylinositol (GPI) anchor.Additionally, many studies have implicated GPI-anchored proteins inproviding specific guidance cues during the outgrowth on neurons inspecific pathways. In studies of the grasshopper nervous system,treatment of embryos with phosphatidylinositol-specific phopholiipase C(PIPLC), which selectively removes GPI-anchored proteins from thesurfaces of cells, resulted in misdirection and faulty navigation amongsubsets of pioneering growth cones, as well as inhibited migratorypatterns of a subset of early neurons, Chang et al., Devel. 114: 507-519(1992). The projection of retinal fibers to the optic tectum appears todepend, in part, on a 33 kDa GPI-anchored protein, however, the precisenature of this protein is unknown. Stahl et al., Neuron 5: 735-743(1990).

[0107] The expression of various GPI-anchored proteins has beencharacterized amongst the different populations of primary rat neuronsamongst dorsal root ganglion, sympathetic neurons of the cervicalganglion, sympathetic neurons of the superior cervical ganglion, andcerebellar granule neurons. Rosen et al., J. Cell Biol. 117: 617-627(1992). In contrast to the similar pattern of total membrane proteinexpression by these different types of neurons, striking differenceswere observed in the expression of GPI-anchored proteins between theseneurons. Recently, a 65 kDa protein band known as neurotrimin wasdiscovered and found to be differentially expressed by primary neurons(Rosen et al., supra), and restricted to the nervous system and found tobe the most abundant and earliest expressed of the GPI-anchored speciesin the CNS. Struyk et al., J. Neuroscience 15(3): 2141-2156 (1995). Thediscovery of neurotrimin has further lead to the identification of afamily of IgSF members, each containing three Ig-like domains that sharesignificant amino acid identity, now termed IgLON. Struyk et al., supra;Pimenta et al., Gene 170(2): 189-95 (1996).

[0108] Additional members of the IgLON subfamily include opiate bindingcell adhesion molecule (OBCAM), Schofield et al., EMBO J. 8: 489-495(1989); limbic associated membrane protein (LAMP), Pimenta et al.,supra; CEPU-1; GP55, Wilson et al., J. Cell Sci. 109: 3129-3138 (1996);Eur. J. Neurosci. 9(2): 334-41 (1997); and AvGp50, Hancox et al., BrainRes. Mol. Brain Res. 44(2): 273-85 (1997).

[0109] While the expression of neurotrimin appears to be widespread, itdoes appear to correlated with the development of several neuralcircuits. For example, between E18 and P10, neurotimin mRNA expressionwithin the forebrain is maintained at high levels in neurons of thedeveloping thalamus, cortical subplate, and cortex, particularly laminaeV and VI (with less intense expression in II, II, and IV, and minimalexpression in lamina I). Cortical subplate neurons may provide an early,temporary scaffold for the ingrowing thalamic afferents en route totheir final synaptic targets in the cortex. Allendoerfer and Shatz,Annu. Rev. Neurosci. 17: 185-218 (1994). Conversely, subplate neuronshave been suggested to be required for cortical neurons from layer V toselect VI to grow into the thalamus, and neurons from layer V to selecttheir targets in the colliculus, pons, and spinal cord (McConnell etal., J. Neurosci. 14: 1892-1907 (1994). The high level expression ofneurotrimin in many of these projections suggests that it could beinvolved in their development.

[0110] In the hindbrain, high levels of neurotrimin message expressionwere observed within the pontine nucleus and by the internal granulecells and Purkinje cells of the cerebellum. The pontine nucleus receivedafferent input from a variety of sources including corticopontine fibersof layer V, and is a major source of afferent input, via mossy fibers,to the granule cells which, in turn, are a major source of afferentinput via parallel fibers to Purkinje cells. [Palay and Chan-Palay, Thecerebellar cortex: cytology and organization. New York: Springer (1974].High level expression of neurotrimin these neurons again suggestspotential involvement in the establishment of these circuits.

[0111] Neurotrimin also exhibits a graded expression pattern in theearly postnatal striatum. Increased neurotrimin expression is foundoverlying the dorsolateral striatum of the rat, while lesserhybridization intensity is seen overlying the ventromedial striatum.Struyk et al., supra. This region of higher neurotrimin hybridizationintensity does not correspond to a cytoarchitecturally differentiableregion, rather it corresponds to the primary area of afferent input fromlayer VI of the contralateral sensorimotor cortex (Gerfen, Nature 311:461-464 (1984); Donoghue and Herkenham, Brain Res. 365: 397-403 (1986)).The ventromedial striatum, by contrast, receives the majority of itsafferent input from the perirhinal and association cortex. It isnoteworthy that a complementary graded pattern of LAMP expression, hasbeen observed within the striatium, with highest expression inventromedial regions, and lowest expression dorsolaterally. Levitt,Science 223: 299-301 (1985); Chesselet et al., Neuroscience 40: 725-733(1991).

[0112] 23. PRO1411

[0113] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many efforts are focused on thescreening of mammalian recombinant DNA libraries to identify the codingsequences for novel secreted proteins. We herein describe theidentification and characterization of a novel secreted proteindesignated herein as PRO1411.

[0114]24. PRO4356

[0115] Glycosylphosphatidylinositol (GPI) anchored proteoglycans aregenerally localized to the cell surface and are thus known to beinvolved in the regulation of responses of cells to numerous growthfactors, cell adhesion molecules and extracellular matrix components.The metastasis-associated GPI-anchored protein (MAGPIAP) is one of thesecell surface proteins which appears to be involved in metastasis.Metastasis is the form of cancer wherein the transformed or malignantcells are traveling and spreading the cancer from one site to another.Therefore, identifying the polypeptides related to metastasis andMAGPIAP is of interest.

[0116] 25. PRO246

[0117] The cell surface protein HCAR is a membrane-bound protein thatacts as a receptor for subgroup C of the adenoviruses and subgroup B ofthe coxsackieviruses. Thus, HCAR may provide a means for mediating viralinfection of cells in that the presence of the HCAR receptor on thecellular surface provides a binding site for viral particles, therebyfacilitating viral infection.

[0118] In light of the physiological importance of membrane-boundproteins and specficially those which serve a cell surface receptor forviruses, efforts are currently being undertaken by both industry andacademia to identify new, native membrane-bound receptor proteins. Manyof these efforts are focused on the screening of mammalian recombinantDNA libraries to identify the coding sequences for novel receptorproteins. We herein describe a novel membrane-bound polypeptide(designated herein as PRO246) having homology to the cell surfaceprotein HCAR and to various tumor antigens including A33 andcarcinoembryonic antigen, wherein this polypeptide may be a novel cellsurface virus receptor or tumor antigen.

[0119] 26. PRO265

[0120] Protein-protein interactions include receptor and antigencomplexes and signaling mechanisms. As more is known about thestructural and functional mechanisms underlying protein-proteininteractions, protein-protein interactions can be more easilymanipulated to regulate the particular result of the protein-proteininteraction. Thus, the underlying mechanisms of protein-proteininteractions are of interest to the scientific and medical community.

[0121] All proteins containing leucine-rich repeats are thought to beinvolved in protein-protein interactions. Leucine-rich repeats are shortsequence motifs present in a number of proteins with diverse functionsand cellular locations. The crystal structure of ribonuclease inhibitorprotein has revealed that leucine-rich repeats correspond to beta-alphastructural units. These units are arranged so that they form a parallelbeta-sheet with one surface exposed to solvent, so that the proteinacquires an unusual, nonglubular shape. These two features have beenindicated as responsible for the protein-binding functions of proteinscontaining leucine-rich repeats. See, Kobe and Deisenhofer, TrendsBiochem. Sci., 19(10):415-421 (October 1994).

[0122] A study has been reported on leucine-rich proteoglycans whichserve as tissue organizers, orienting and ordering collagen fibrilsduring ontogeny and are involved in pathological processes such as woundhealing, tissue repair, and tumor stroma formation. lozzo, R. V., Crit.Rev. Biochem. Mol. Biol., 32(2):141-174 (1997). Others studiesimplicating leucine rich proteins in wound healing and tissue repair areDe La Salle, C., et al., Vouv. Rev. Fr. Hematol. (Germany),37(4):215-222 (1995), reporting mutations in the leucine rich motif in acomplex associated with the bleeding disorder Bernard-Soulier syndromeand Chlemetson, K. J., Thromb. Haemost. (Germany), 74(1):111-116 (July1995), reporting that platelets have leucine rich repeats. Anotherprotein of particular interest which has been reported to haveleucine-rich repeats is the SLIT protein which has been reported to beuseful in treating neuro-degenerative diseases such as Alzheimer'sdisease, nerve damage such as in Parkinson's disease, and for diagnosisof cancer, see, Artavanistsakonas, S. and Rothberg, J. M., WO9210518-A1by Yale University. Other studies reporting on the biological functionsof proteins having leucine-rich repeats include: Tayar, N., et al., Mol.Cell Endocrinol., (Ireland), 125(1-2):65-70 (December 1996)(gonadotropin receptor involvement); Miura, Y., et al., Nippon Rinsho(Japan), 54(7):1784-1789 (July 1996) (apoptosis involvement); Harris, P.C., et al., J. Am. Soc. Nephrol., 6(4):1125-1133 (October 1995) (kidneydisease involvement); and Ruoslahti, E. I., et al., WO9110727-A by LaJolla Cancer Research Foundation (decorin binding to transforming growthfactor-β involvement for treatment for cancer, wound healing andscarring). Also of particular interest is fibromodulin and its use toprevent or reduce dermal scarring. A study of fibromodulin is found inU.S. Pat. No. 5,654,270 to Ruoslahti, et al.

[0123] Efforts are therefore being undertaken by both industry andacademia to identify new proteins having leucine rich repeats to betterunderstand protein-protein interactions. Of particular interest arethose proteins having leucine rich repeats and homology to knownproteins having leucine rich repeats such as fibromodulin, the SLITprotein and platelet glycoprotein V. Many efforts are focused on thescreening of mammalian recombinant DNA libraries to identify the codingsequences for novel secreted and membrane-bound proteins having leucinerich repeats. We herein describe the identification and characterizationof novel polypeptides having homology to fibromodulin, herein designatedas PRO265 polypeptides.

[0124] 27. PRO941

[0125] Cadherins are a large family of transmembrane proteins. Cadherinscomprise a family of calcium-dependent glycoproteins that function inmediating cell-cell adhesion in virtually all solid tissues ofmulticellular organisms. At least cadherins 1-13 as well as types B, E,EP, M, N, P and R have been identified and characterized. Among thefunctions cadherins are known for, with some exceptions, are thatcadherins participate in cell aggregation and are associated withcell-cell adhesion sites. Recently, it has been reported that while allcadherins share multiple repeats of a cadherin specific motif believedto correspond to folding of extracellular domains, members of thecadherin superfamily have divergent structures and, possibly, functions.In particular it has been reported that members of the cadherinsuperfamily are involved in signal transduction. See, Suzuki, J. CellBiochem., 61(4):531-542 (1996). Cadherins are further described inTanihara et al., J. Cell Sci., 107(6):1697-1704 (1994), Aberle et al.,J. Cell Biochem., 61(4):514-523 (1996) and Tanihara et al., Cell Adhes.Commun., 2(1):15-26 (1994). We herein describe the identification andcharacterization of a novel polypeptide having homology to a cadherinprotein, designated herein as PRO941.

[0126]28. PRO10096

[0127] Interleukin-10 (IL-10) is a pleiotropic immunosuppressivecytokine that has been implicated as an important regulator of thefunctions of myeloid and lymphoid cells. It has been demonstrated thatIL-10 functions as a potent inhibitor of the activation of the synthesisof various inflammatory cytokines including, for example, IL-1, IL-6,IFN-γ and TNF-α (Gesser et al., Proc. Natl. Acad. Sci. USA94:14620-14625 (1997)). Moreover, IL-10 has been demonstrated tostrongly inhibit several of the accessory activities of macrophages,thereby functioning as a potent suppressor of the effector functions ofmacrophages, T-cells and NK cells (Kuhn et al., Cell 75:263-274 (1993)).Furthermore, IL-10 has been strongly implicated in the regulation ofB-cell, mast cell and thymocyte differentiation.

[0128] IL-10 was independently identified in two separate lines ofexperiments. First, cDNA clones encoding murine IL-10 were identifiedbased upon the expression of cytokine synthesis inhibitory factor (Mooreet al., Science 248:1230-1234 (1990)), wherein the human IL-10counterpart cDNAs were subsequently identified by cross-hybridizationwith the murine IL-10 cDNA (Viera et al., Proc. Natl. Acad. Sci. USA88:1172-1176 (1991)). Additionally, IL-10 was independently identifiedas a B-cell-derived mediator which functioned to co-stimulate activethymocytes (Suda et al., Cell Immunol. 129:228 (1990)).

[0129] We herein describe the identification and characterization ofnovel polypeptides having sequence similarity to IL-10, designatedherein as PRO10096 polypeptides.

[0130] 29. PRO6003

[0131] Efforts are being undertaken by both industry and academia toidentify new, native receptor or membrane-bound proteins. Many effortsare focused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel receptor or membrane-boundproteins. We herein describe the identification and characterization ofnovel polypeptides designated herein as PRO6003 polypeptides.

SUMMARY OF THE INVENTION

[0132] In one embodiments of the present invention, the inventionprovides vectors comprising DNA encoding any of the herein describedpolypeptides. Host cell comprising any such vector are also provided. Byway of example, the host cells may be CHO cells, E. coli, or yeast. Aprocess for producing any of the herein described polypeptides isfurther provided and comprises culturing host cells under conditionssuitable for expression of the desired polypeptide and recovering thedesired polypeptide from the cell culture.

[0133] In other embodiments, the invention provides chimeric moleculescomprising any of the herein described polypeptides fused to aheterologous polypeptide or amino acid sequence. Example of suchchimeric molecules comprise any of the herein described polypeptidesfused to an epitope tag sequence or a Fc region of an immunoglobulin.

[0134] In another embodiment, the invention provides an antibody whichspecifically binds to any of the above or below described polypeptides.Optionally, the antibody is a monoclonal antibody, humanized antibody,antibody fragment or single-chain antibody.

[0135] In yet other embodiments, the invention provides oligonucleotideprobes useful for isolating genomic and cDNA nucleotide sequences or asantisense probes, wherein those probes may be derived from any of theabove or below described nucleotide sequences.

[0136] In other embodiments, the invention provides an isolated nucleicacid molecule comprising a nucleotide sequence that encodes a PROpolypeptide.

[0137] In one aspect, the isolated nucleic acid molecule comprises anucleotide sequence having at least about 80% nucleic acid sequenceidentity, alternatively at least about 81% nucleic acid sequenceidentity, alternatively at least about 82% nucleic acid sequenceidentity, alternatively at least about 83% nucleic acid sequenceidentity, alternatively at least about 84% nucleic acid sequenceidentity, alternatively at least about 85% nucleic acid sequenceidentity, alternatively at least about 86% nucleic acid sequenceidentity, alternatively at least about 87% nucleic acid sequenceidentity, alternatively at least about 88% nucleic acid sequenceidentity, alternatively at least about 89% nucleic acid sequenceidentity, alternatively at least about 90% nucleic acid sequenceidentity, alternatively at least about 91% nucleic acid sequenceidentity, alternatively at least about 92% nucleic acid sequenceidentity, alternatively at least about 93% nucleic acid sequenceidentity, alternatively at least about 94% nucleic acid sequenceidentity, alternatively at least about 95% nucleic acid sequenceidentity, alternatively at least about 96% nucleic acid sequenceidentity, alternatively at least about 97% nucleic acid sequenceidentity, alternatively at least about 98% nucleic acid sequenceidentity and alternatively at least about 99% nucleic acid sequenceidentity to (a) a DNA molecule encoding a PRO polypeptide having afull-length amino acid sequence as disclosed herein, an amino acidsequence lacking the signal peptide as disclosed herein, anextracellular domain of a transmembrane protein, with or without thesignal peptide, as disclosed herein or any other specifically definedfragment of the full-length amino acid sequence as disclosed herein, or(b) the complement of the DNA molecule of (a).

[0138] In other aspects, the isolated nucleic acid molecule comprises anucleotide sequence having at least about 80% nucleic acid sequenceidentity, alternatively at least about 81% nucleic acid sequenceidentity, alternatively at least about 82% nucleic acid sequenceidentity, alternatively at least about 83% nucleic acid sequenceidentity, alternatively at least about 84% nucleic acid sequenceidentity, alternatively at least about 85% nucleic acid sequenceidentity, alternatively at least about 86% nucleic acid sequenceidentity, alternatively at least about 87% nucleic acid sequenceidentity, alternatively at least about 88% nucleic acid sequenceidentity, alternatively at least about 89% nucleic acid sequenceidentity, alternatively at least about 90% nucleic acid sequenceidentity, alternatively at least about 91% nucleic acid sequenceidentity, alternatively at least about 92% nucleic acid sequenceidentity, alternatively at least about 93% nucleic acid sequenceidentity, alternatively at least about 94% nucleic acid sequenceidentity, alternatively at least about 95% nucleic acid sequenceidentity, alternatively at least about 96% nucleic acid sequenceidentity, alternatively at least about 97% nucleic acid sequenceidentity, alternatively at least about 98% nucleic acid sequenceidentity and alternatively at least about 99% nucleic acid sequenceidentity to (a) a DNA molecule comprising the coding sequence of afull-length PRO polypeptide cDNA as disclosed herein, the codingsequence of a PRO polypeptide lacking the signal peptide as disclosedherein, the coding sequence of an extracellular domain of atransmembrane PRO polypeptide, with or without the signal peptide, asdisclosed herein or the coding sequence of any other specificallydefined fragment of the full-length amino acid sequence as disclosedherein, or (b) the complement of the DNA molecule of (a).

[0139] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising a nucleotide sequence having at least about 80%nucleic acid sequence identity, alternatively at least about 81% nucleicacid sequence identity, alternatively at least about 82% nucleic acidsequence identity, alternatively at least about 83% nucleic acidsequence identity, alternatively at least about 84% nucleic acidsequence identity, alternatively at least about 85% nucleic acidsequence identity, alternatively at least about 86% nucleic acidsequence identity, alternatively at least about 87% nucleic acidsequence identity, alternatively at least about 88% nucleic acidsequence identity, alternatively at least about 89% nucleic acidsequence identity, alternatively at least about 90% nucleic acidsequence identity, alternatively at least about 91% nucleic acidsequence identity, alternatively at least about 92% nucleic acidsequence identity, alternatively at least about 93% nucleic acidsequence identity, alternatively at least about 94% nucleic acidsequence identity, alternatively at least about 95% nucleic acidsequence identity, alternatively at least about 96% nucleic acidsequence identity, alternatively at least about 97% nucleic acidsequence identity, alternatively at least about 98% nucleic acidsequence identity and alternatively at least about 99% nucleic acidsequence identity to (a) a DNA molecule that encodes the same maturepolypeptide encoded by any of the human protein cDNAs deposited with theATCC as disclosed herein, or (b) the complement of the DNA molecule of(a).

[0140] Another aspect the invention provides an isolated nucleic acidmolecule comprising a nucleotide sequence encoding a PRO polypeptidewhich is either transmembrane domain-deleted or transmembranedomain-inactivated, or is complementary to such encoding nucleotidesequence, wherein the transmembrane domain(s) of such polypeptide aredisclosed herein. Therefore, soluble extracellular domains of the hereindescribed PRO polypeptides are contemplated.

[0141] Another embodiment is directed to fragments of a PRO polypeptidecoding sequence, or the complement thereof, that may find use as, forexample, hybridization probes, for encoding fragments of a PROpolypeptide that may optionally encode a polypeptide comprising abinding site for an anti-PRO antibody or as antisense oligonucleotideprobes. Such nucleic acid fragments are usually at least about 20nucleotides in length, alternatively at least about 30 nucleotides inlength, alternatively at least about 40 nucleotides in length,alternatively at least about 50 nucleotides in length, alternatively atleast about 60 nucleotides in length, alternatively at least about 70nucleotides in length, alternatively at least about 80 nucleotides inlength, alternatively at least about 90 nucleotides in length,alternatively at least about 100 nucleotides in length, alternatively atleast about 110 nucleotides in length, alternatively at least about 120nucleotides in length, alternatively at least about 130 nucleotides inlength, alternatively at least about 140 nucleotides in length,alternatively at least about 150 nucleotides in length, alternatively atleast about 160 nucleotides in length, alternatively at least about 170nucleotides in length, alternatively at least about 180 nucleotides inlength, alternatively at least about 190 nucleotides in length,alternatively at least about 200 nucleotides in length, alternatively atleast about 250 nucleotides in length, alternatively at least about 300nucleotides in length, alternatively at least about 350 nucleotides inlength, alternatively at least about 400 nucleotides in length,alternatively at least about 450 nucleotides in length, alternatively atleast about 500 nucleotides in length, alternatively at least about 600nucleotides in length, alternatively at least about 700 nucleotides inlength, alternatively at least about 800 nucleotides in length,alternatively at least about 900 nucleotides in length and alternativelyat least about 1000 nucleotides in length, wherein in this context theterm “about” means the referenced nucleotide sequence length plus orminus 10% of that referenced length. It is noted that novel fragments ofa PRO polypeptide-encoding nucleotide sequence may be determined in aroutine manner by aligning the PRO polypeptide-encoding nucleotidesequence with other known nucleotide sequences using any of a number ofwell known sequence alignment programs and determining which PROpolypeptide-encoding nucleotide sequence fragment(s) are novel. All ofsuch PRO polypeptide-encoding nucleotide sequences are contemplatedherein. Also contemplated are the PRO polypeptide fragments encoded bythese nucleotide molecule fragments, preferably those PRO polypeptidefragments that comprise a binding site for an anti-PRO antibody.

[0142] In another embodiment, the invention provides isolated PROpolypeptide encoded by any of the isolated nucleic acid sequenceshereinabove identified.

[0143] In a certain aspect, the invention concerns an isolated PROpolypeptide, comprising an amino acid sequence having at least about 80%amino acid sequence identity, alternatively at least about 81% aminoacid sequence identity, alternatively at least about 82% amino acidsequence identity, alternatively at least about 83% amino acid sequenceidentity, alternatively at least about 84% amino acid sequence identity,alternatively at least about 85% amino acid sequence identity,alternatively at least about 86% amino acid sequence identity,alternatively at least about 87% amino acid sequence identity,alternatively at least about 88% amino acid sequence identity,alternatively at least about 89% amino acid sequence identity,alternatively at least about 90% amino acid sequence identity,alternatively at least about 91% amino acid sequence identity,alternatively at least about 92% amino acid sequence identity,alternatively at least about 93% amino acid sequence identity,alternatively at least about 94% amino acid sequence identity,alternatively at least about 95% amino acid sequence identity,alternatively at least about 96% amino acid sequence identity,alternatively at least about 97% amino acid sequence identity,alternatively at least about 98% amino acid sequence identity andalternatively at least about 99% amino acid sequence identity to a PROpolypeptide having a full-length amino acid sequence as disclosedherein, an amino acid sequence lacking the signal peptide as disclosedherein, an extracellular domain of a transmembrane protein, with orwithout the signal peptide, as disclosed herein or any otherspecifically defined fragment of the full-length amino acid sequence asdisclosed herein.

[0144] In a further aspect, the invention concerns an isolated PROpolypeptide comprising an amino acid sequence having at least about 80%amino acid sequence identity, alternatively at least about 81% aminoacid sequence identity, alternatively at least about 82% amino acidsequence identity, alternatively at least about 83% amino acid sequenceidentity, alternatively at least about 84% amino acid sequence identity,alternatively at least about 85% amino acid sequence identity,alternatively at least about 86% amino acid sequence identity,alternatively at least about 87% amino acid sequence identity,alternatively at least about 88% amino acid sequence identity,alternatively at least about 89% amino acid sequence identity,alternatively at least about 90% amino acid sequence identity,alternatively at least about 91% amino acid sequence identity,alternatively at least about 92% amino acid sequence identity,alternatively at least about 93% amino acid sequence identity,alternatively at least about 94% amino acid sequence identity,alternatively at least about 95% amino acid sequence identity,alternatively at least about 96% amino acid sequence identity,alternatively at least about 97% amino acid sequence identity,alternatively at least about 98% amino acid sequence identity andalternatively at least about 99% amino acid sequence identity to anamino acid sequence encoded by any of the human protein cDNAs depositedwith the ATCC as disclosed herein.

[0145] In a further aspect, the invention concerns an isolated PROpolypeptide comprising an amino acid sequence scoring at least about 80%positives, alternatively at least about 81% positives, alternatively atleast about 82% positives, alternatively at least about 83% positives,alternatively at least about 84% positives, alternatively at least about85% positives, alternatively at least about 86% positives, alternativelyat least about 87% positives, alternatively at least about 88%positives, alternatively at least about 89% positives, alternatively atleast about 90% positives, alternatively at least about 91% positives,alternatively at least about 92% positives, alternatively at least about93% positives, alternatively at least about 94% positives, alternativelyat least about 95% positives, alternatively at least about 96%positives, alternatively at least about 97% positives, alternatively atleast about 98% positives and alternatively at least about 99% positiveswhen compared with the amino acid sequence of a PRO polypeptide having afull-length amino acid sequence as disclosed herein, an amino acidsequence lacking the signal peptide as disclosed herein, anextracellular domain of a transmembrane protein, with or without thesignal peptide, as disclosed herein or any other specifically definedfragment of the full-length amino acid sequence as disclosed herein.

[0146] In a specific aspect, the invention provides an isolated PROpolypeptide without the N-terminal signal sequence and/or the initiatingmethionine and is encoded by a nucleotide sequence that encodes such anamino acid sequence as hereinbefore described. Processes for producingthe same are also herein described, wherein those processes compriseculturing a host cell comprising a vector which comprises theappropriate encoding nucleic acid molecule under conditions suitable forexpression of the PRO polypeptide and recovering the PRO polypeptidefrom the cell culture.

[0147] Another aspect the invention provides an isolated PRO polypeptidewhich is either transmembrane domain-deleted or transmembranedomain-inactivated. Processes for producing the same are also hereindescribed, wherein those processes comprise culturing a host cellcomprising a vector which comprises the appropriate encoding nucleicacid molecule under conditions suitable for expression of the PROpolypeptide and recovering the PRO polypeptide from the cell culture.

[0148] In yet another embodiment, the invention concerns agonists andantagonists of a native PRO polypeptide as defined herein. In aparticular embodiment, the agonist or antagonist is an anti-PRO antibodyor a small molecule.

[0149] In a further embodiment, the invention concerns a method ofidentifying agonists or antagonists to a PRO polypeptide which comprisecontacting the PRO polypeptide with a candidate molecule and monitoringa biological activity mediated by said PRO polypeptide. Preferably, thePRO polypeptide is a native PRO polypeptide.

[0150] In a still further embodiment, the invention concerns acomposition of matter comprising a PRO polypeptide, or an agonist orantagonist of a PRO polypeptide as herein described, or an anti-PROantibody, in combination with a carrier. Optionally, the carrier is apharmaceutically acceptable carrier.

[0151] Another embodiment of the present invention is directed to theuse of a PRO polypeptide, or an agonist or antagonist thereof ashereinbefore described, or an anti-PRO antibody, for the preparation ofa medicament useful in the treatment of a condition which is responsiveto the PRO polypeptide, an agonist or antagonist thereof or an anti-PROantibody.

BRIEF DESCRIPTION OF THE DRAWINGS

[0152]FIG. 1 shows a nucleotide sequence (SEQ ID NO: 3) of a nativesequence PRO196 cDNA, wherein SEQ ID NO: 3 is a clone designated hereinas “DNA22779-1130”.

[0153]FIG. 2 shows the amino acid sequence (SEQ ID NO: 4) derived fromthe coding sequence of SEQ ID NO: 3 shown in FIG. 1.

[0154]FIG. 3 shows a nucleotide sequence (SEQ ID NO: 8) of a nativesequence PRO444 cDNA, wherein SEQ ID NO: 8 is a clone designated hereinas “DNA26846-1397”.

[0155]FIG. 4 shows the amino acid sequence (SEQ ID NO: 9) derived fromthe coding sequence of SEQ ID NO: 8 shown in FIG. 3.

[0156]FIG. 5 shows a nucleotide sequence (SEQ ID NO: 10) of a nativesequence PRO183 cDNA, wherein SEQ ID NO: 10 is a clone designated hereinas “DNA28498”.

[0157]FIG. 6 shows the amino acid sequence (SEQ ID NO: 11) derived fromthe coding sequence of SEQ ID NO: 10 shown in FIG. 5.

[0158]FIG. 7 shows a nucleotide sequence (SEQ ID NO: 12) of a nativesequence PRO 185 cDNA, wherein SEQ ID NO: 12 is a clone designatedherein as “DNA28503”.

[0159]FIG. 8 shows the amino acid sequence (SEQ ID NO: 13) derived fromthe coding sequence of SEQ ID NO: 12 shown in FIG. 7.

[0160]FIG. 9 shows a nucleotide sequence (SEQ ID NO: 14) of a nativesequence PRO210 cDNA, wherein SEQ ID NO: 14 is a clone designated hereinas “DNA32279-1131”.

[0161]FIG. 10 shows the amino acid sequence (SEQ ID NO: 15) derived fromthe coding sequence of SEQ ID NO: 14 shown in FIG. 9.

[0162]FIG. 11 shows a nucleotide sequence (SEQ ID NO: 16) of a nativesequence PRO215 cDNA, wherein SEQ ID NO: 16 is a clone designated hereinas “DNA32288-1132”.

[0163]FIG. 12 shows the amino acid sequence (SEQ ID NO: 17) derived fromthe coding sequence of SEQ ID NO: 16 shown in FIG. 11.

[0164]FIG. 13 shows a nucleotide sequence (SEQ ID NO: 21) of a nativesequence PRO217 cDNA, wherein SEQ ID NO: 21 is a clone designated hereinas “DNA33094-1131”.

[0165]FIG. 14 shows the amino acid sequence (SEQ ID NO: 22) derived fromthe coding sequence of SEQ ID NO: 21 shown in FIG. 13.

[0166]FIG. 15 shows a nucleotide sequence (SEQ ID NO: 23) of a nativesequence PRO242 cDNA, wherein SEQ ID NO: 23 is a clone designated hereinas “DNA33785-1143”.

[0167]FIG. 16 shows the amino acid sequence (SEQ ID NO: 24) derived fromthe coding sequence of SEQ ID NO: 23 shown in FIG. 15.

[0168]FIG. 17 shows a nucleotide sequence (SEQ ID NO: 28) of a nativesequence PRO288 cDNA, wherein SEQ ID NO: 28 is a clone designated hereinas “DNA35663-1129”.

[0169]FIG. 18 shows the amino acid sequence (SEQ ID NO: 29) derived fromthe coding sequence of SEQ ID NO: 28 shown in FIG. 17.

[0170]FIG. 19 shows a nucleotide sequence (SEQ ID NO: 31) of a nativesequence PRO365 cDNA, wherein SEQ ID NO: 31 is a clone designated hereinas “DNA46777-1253”.

[0171]FIG. 20 shows the amino acid sequence (SEQ ID NO: 32) derived fromthe coding sequence of SEQ ID NO: 31 shown in FIG. 19.

[0172]FIG. 21 shows a nucleotide sequence (SEQ ID NO: 38) of a nativesequence PRO1361 cDNA, wherein SEQ ID NO: 38 is a clone designatedherein as “DNA60783-1611”.

[0173]FIG. 22 shows the amino acid sequence (SEQ ID NO: 39) derived fromthe coding sequence of SEQ ID NO: 38 shown in FIG. 21.

[0174]FIG. 23 shows a nucleotide sequence (SEQ ID NO: 40) of a nativesequence PRO1308 cDNA, wherein SEQ ID NO: 40 is a clone designatedherein as “DNA62306-1570”.

[0175]FIG. 24 shows the amino acid sequence (SEQ ID NO: 41) derived fromthe coding sequence of SEQ ID NO: 40 shown in FIG. 23.

[0176]FIG. 25 shows a nucleotide sequence (SEQ ID NO: 51) of a nativesequence PRO1183 cDNA, wherein SEQ ID NO: 51 is a clone designatedherein as “DNA62880-1513”.

[0177]FIG. 26 shows the amino acid sequence (SEQ ID NO: 52) derived fromthe coding sequence of SEQ ID NO: 51 shown in FIG. 25.

[0178]FIG. 27 shows a nucleotide sequence (SEQ ID NO: 53) of a nativesequence PRO1272 cDNA, wherein SEQ ID NO: 53 is a clone designatedherein as “DNA64896-1539”.

[0179]FIG. 28 shows the amino acid sequence (SEQ ID NO: 54) derived fromthe coding sequence of SEQ ID NO: 53 shown in FIG. 27.

[0180]FIG. 29 shows a nucleotide sequence (SEQ ID NO: 55) of a nativesequence PRO1419 cDNA, wherein SEQ ID NO: 55 is a clone designatedherein as “DNA71290-1630”.

[0181]FIG. 30 shows the amino acid sequence (SEQ ID NO: 56) derived fromthe coding sequence of SEQ ID NO: 55 shown in FIG. 29.

[0182]FIG. 31 shows a nucleotide sequence (SEQ ID NO: 57) of a nativesequence PRO4999 cDNA, wherein SEQ ID NO: 57 is a clone designatedherein as “DNA96031-2664”.

[0183]FIG. 32 shows the amino acid sequence (SEQ ID NO: 58) derived fromthe coding sequence of SEQ ID NO: 57 shown in FIG. 31.

[0184]FIG. 33 shows a nucleotide sequence (SEQ ID NO: 62) of a nativesequence PRO7170 cDNA, wherein SEQ ID NO: 62 is a clone designatedherein as “DNA108722-2743”.

[0185]FIG. 34 shows the amino acid sequence (SEQ ID NO: 63) derived fromthe coding sequence of SEQ ID NO: 62 shown in FIG. 33.

[0186]FIG. 35 shows a nucleotide sequence (SEQ ID NO: 64) of a nativesequence PRO248 cDNA, wherein SEQ ID NO: 64 is a clone designated hereinas “DNA35674-1142”.

[0187]FIG. 36 shows the amino acid sequence (SEQ ID NO: 65) derived fromthe coding sequence of SEQ ID NO: 64 shown in FIG. 35.

[0188]FIG. 37 shows a nucleotide sequence (SEQ ID NO: 72) of a nativesequence PRO353 cDNA, wherein SEQ ID NO: 72 is a clone designated hereinas “DNA41234”.

[0189]FIG. 38 shows the amino acid sequence (SEQ ID NO: 73) derived fromthe coding sequence of SEQ ID NO: 72 shown in FIG. 37.

[0190]FIG. 39 shows a nucleotide sequence (SEQ ID NO: 77) of a nativesequence PRO1318 cDNA, wherein SEQ ID NO: 77 is a clone designatedherein as “DNA73838-1674”.

[0191]FIG. 40 shows the amino acid sequence (SEQ ID NO: 78) derived fromthe coding sequence of SEQ ID NO: 77 shown in FIG. 39.

[0192]FIG. 41 shows a nucleotide sequence (SEQ ID NO: 79) of a nativesequence PRO1600 cDNA, wherein SEQ ID NO: 79 is a clone designatedherein as “DNA77503-1686”.

[0193]FIG. 42 shows the amino acid sequence (SEQ ID NO: 80) derived fromthe coding sequence of SEQ ID NO: 79 shown in FIG. 41.

[0194]FIG. 43 shows a nucleotide sequence (SEQ ID NO: 83) of a nativesequence PRO9940 cDNA, wherein SEQ ID NO: 83 is a clone designatedherein as “DNA92282”.

[0195]FIG. 44 shows the amino acid sequence (SEQ ID NO: 84) derived fromthe coding sequence of SEQ ID NO: 83 shown in FIG. 43.

[0196]FIG. 45 shows a nucleotide sequence (SEQ ID NO: 85) of a nativesequence PRO533 cDNA, wherein SEQ ID NO: 85 is a clone designated hereinas “DNA49435-1219”.

[0197]FIG. 46 shows the amino acid sequence (SEQ ID NO: 86) derived fromthe coding sequence of SEQ ID NO: 85 shown in FIG. 45.

[0198]FIG. 47 shows a nucleotide sequence (SEQ ID NO: 90) of a nativesequence PRO301 cDNA, wherein SEQ ID NO: 90 is a clone designated hereinas “DNA40628-1216”.

[0199]FIG. 48 shows the amino acid sequence (SEQ ID NO: 91) derived fromthe coding sequence of SEQ ID NO: 90 shown in FIG. 47.

[0200]FIG. 49 shows a nucleotide sequence (SEQ ID NO: 98) of a nativesequence PRO187 cDNA, wherein SEQ ID NO: 98 is a clone designated hereinas “DNA27864-1155”.

[0201]FIG. 50 shows the amino acid sequence (SEQ ID NO: 99) derived fromthe coding sequence of SEQ ID NO: 98 shown in FIG. 49.

[0202]FIG. 51 shows a nucleotide sequence (SEQ ID NO: 103) of a nativesequence PRO337 cDNA, wherein SEQ ID NO: 103 is a clone designatedherein as “DNA43316-1237”.

[0203]FIG. 52 shows the amino acid sequence (SEQ ID NO: 104) derivedfrom the coding sequence of SEQ ID NO: 103 shown in FIG. 51.

[0204]FIG. 53 shows a nucleotide sequence (SEQ ID NO: 105) of a nativesequence PRO141 1 cDNA, wherein SEQ ID NO: 105 is a clone designatedherein as “DNA59212-1627”.

[0205]FIG. 54 shows the amino acid sequence (SEQ ID NO: 106) derivedfrom the coding sequence of SEQ ID NO: 105 shown in FIG. 53.

[0206]FIG. 55 shows a nucleotide sequence (SEQ ID NO: 107) of a nativesequence PRO4356 cDNA, wherein SEQ ID NO: 107 is a clone designatedherein as “DNA86576-2595”.

[0207]FIG. 56 shows the amino acid sequence (SEQ ID NO: 108) derivedfrom the coding sequence of SEQ ID NO: 107 shown in FIG. 55.

[0208]FIG. 57 shows a nucleotide sequence (SEQ ID NO: 109) of a nativesequence PRO246 cDNA, wherein SEQ ID NO: 109 is a clone designatedherein as “DNA35639-1172”.

[0209]FIG. 58 shows the amino acid sequence (SEQ ID NO: 110) derivedfrom the coding sequence of SEQ ID NO: 109 shown in FIG. 57.

[0210]FIG. 59 shows a nucleotide sequence (SEQ ID NO: 114) of a nativesequence PRO265 cDNA, wherein SEQ ID NO: 114 is a clone designatedherein as “DNA36350-1158”.

[0211]FIG. 60 shows the amino acid sequence (SEQ ID NO: 115) derivedfrom the coding sequence of SEQ ID NO: 114 shown in FIG. 59.

[0212]FIG. 61 shows a nucleotide sequence (SEQ ID NO: 120) of a nativesequence PRO941 cDNA, wherein SEQ ID NO: 120 is a clone designatedherein as “DNA53906-1368”.

[0213]FIG. 62 shows the amino acid sequence (SEQ ID NO: 121) derivedfrom the coding sequence of SEQ ID NO: 120 shown in FIG. 61.

[0214]FIG. 63 shows a nucleotide sequence (SEQ ID NO: 125) of a nativesequence PRO10096 cDNA, wherein SEQ ID NO: 125 is a clone designatedherein as “DNA125185-2806”.

[0215]FIG. 64 shows the amino acid sequence (SEQ ID NO: 126) derivedfrom the coding sequence of SEQ ID NO: 125 shown in FIG. 63.

[0216]FIG. 65 shows a nucleotide sequence (SEQ ID NO: 127) of a nativesequence PRO6003 cDNA, wherein SEQ ID NO: 127 is a clone designatedherein as “DNA83568-2692”.

[0217]FIG. 66 shows the amino acid sequence (SEQ ID NO: 128) derivedfrom the coding sequence of SEQ ID NO: 127 shown in FIG. 65.

[0218] FIGS. 67A-B show a nucleotide sequence (SEQ ID NO: 129) of anative sequence PRO6004 cDNA, wherein SEQ ID NO: 129 is a clonedesignated herein as “DNA92259”.

[0219]FIG. 68 shows the amino acid sequence (SEQ ID NO: 130) derivedfrom the coding sequence of SEQ ID NO: 129 shown in FIGS. 67A-B.

[0220]FIG. 69 shows a nucleotide sequence (SEQ ID NO: 131) of a nativesequence PRO350 cDNA, wherein SEQ ID NO: 131 is a clone designatedherein as “DNA44175-1314”.

[0221]FIG. 70 shows the amino acid sequence (SEQ ID NO: 132) derivedfrom the coding sequence of SEQ ID NO: 131 shown in FIG. 69.

[0222]FIG. 71 shows a nucleotide sequence (SEQ ID NO: 136) of a nativesequence PRO2630 cDNA, wherein SEQ ID NO: 136 is a clone designatedherein as “DNA83551”.

[0223]FIG. 72 shows the amino acid sequence (SEQ ID NO: 137) derivedfrom the coding sequence of SEQ ID NO: 136 shown in FIG. 71.

[0224]FIG. 73 shows a nucleotide sequence (SEQ ID NO: 138) of a nativesequence PRO6309 cDNA, wherein SEQ ID NO: 138 is a clone designatedherein as “DNA1 16510”.

[0225]FIG. 74 shows the amino acid sequence (SEQ ID NO: 139) derivedfrom the coding sequence of SEQ ID NO: 138 shown in FIG. 73.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0226] 1. Definitions

[0227] The terms “PRO polypeptide” and “PRO” as used herein and whenimmediately followed by a numerical designation refer to variouspolypeptides, wherein the complete designation (i.e., PRO/number) refersto specific polypeptide sequences as described herein. The terms“PRO/number polypeptide” and “PRO/number” wherein the term “number” isprovided as an actual numerical designation as used herein encompassnative sequence polypeptides and polypeptide variants (which are furtherdefined herein). The PRO polypeptides described herein may be isolatedfrom a variety of sources, such as from human tissue types or fromanother source, or prepared by recombinant or synthetic methods. Theterm “PRO polypeptide” refers to each individual PRO/number polypeptidedisclosed herein. All disclosures in this specification which refer tothe “PRO polypeptide” refer to each of the polypeptides individually aswell as jointly. For example, descriptions of the preparation of,purification of, derivation of, formation of antibodies to or against,administration of, compositions containing, treatment of a disease with,etc., pertain to each polypeptide of the invention individually. Theterm “PRO polypeptide” also includes variants of the PRO/numberpolypeptides disclosed herein.

[0228] A “native sequence PRO polypeptide” comprises a polypeptidehaving the same amino acid sequence as the corresponding PRO polypeptidederived from nature. Such native sequence PRO polypeptides can beisolated from nature or can be produced by recombinant or syntheticmeans. The term “native sequence PRO polypeptide” specificallyencompasses naturally-occurring truncated or secreted forms of thespecific PRO polypeptide (e.g., an extracellular domain sequence),naturally-occurring variant forms (e.g., alternatively spliced forms)and naturally-occurring allelic variants of the polypeptide. In variousembodiments of the invention, the native sequence PRO polypeptidesdisclosed herein are mature or full-length native sequence polypeptidescomprising the full-length amino acids sequences shown in theaccompanying figures. Start and stop codons are shown in bold font andunderlined in the figures. However, while the PRO polypeptide disclosedin the accompanying figures are shown to begin with methionine residuesdesignated herein as amino acid position 1 in the figures, it isconceivable and possible that other methionine residues located eitherupstream or downstream from the amino acid position 1 in the figures maybe employed as the starting amino acid residue for the PRO polypeptides.

[0229] The PRO polypeptide “extracellular domain” or “ECD” refers to aform of the. PRO polypeptide which is essentially free of thetransmembrane and cytoplasmic domains. Ordinarily, a PRO polypeptide ECDwill have less than 1% of such transmembrane and/or cytoplasmic domainsand preferably, will have less than 0.5% of such domains. It will beunderstood that any transmembrane domains identified for the PROpolypeptides of the present invention are identified pursuant tocriteria routinely employed in the art for identifying that type ofhydrophobic domain. The exact boundaries of a transmembrane domain mayvary but most likely by no more than about 5 amino acids at either endof the domain as initially identified herein. Optionally, therefore, anextracellular domain of a PRO polypeptide may contain from about 5 orfewer amino acids on either side of the transmembranedomain/extracellular domain boundary as identified in the Examples orspecification and such polypeptides, with or without the associatedsignal peptide, and nucleic acid encoding them, are comtemplated by thepresent invention.

[0230] The approximate location of the “signal peptides” of the variousPRO polypeptides disclosed herein are shown in the present specificationand/or the accompanying figures. It is noted, however, that theC-terminal boundary of a signal peptide may vary, but most likely by nomore than about 5 amino acids on either side of the signal peptideC-terminal boundary as initially identified herein, wherein theC-terminal boundary of the signal peptide may be identified pursuant tocriteria routinely employed in the art for identifying that type ofamino acid sequence element (e.g., Nielsen et al., Prot. Eng. 10: 1-6(1997) and von Heinje et al., Nucl. Acids. Res. 14:4683-4690 (1986)).Moreover, it is also recognized that, in some cases, cleavage of asignal sequence from a secreted polypeptide is not entirely uniform,resulting in more than one secreted species. These mature polypeptides,where the signal peptide is cleaved within no more than about 5 aminoacids on either side of the C-terminal boundary of the signal peptide asidentified herein, and the polynucleotides encoding them, arecontemplated by the present invention.

[0231] “PRO polypeptide variant” means an active PRO polypeptide asdefined above or below having at least about 80% amino acid sequenceidentity with a full-length native sequence PRO polypeptide sequence asdisclosed herein, a PRO polypeptide sequence lacking the signal peptideas disclosed herein, an extracellular domain of a PRO polypeptide, withor without the signal peptide, as disclosed herein or any other fragmentof a full-length PRO polypeptide sequence as disclosed herein. Such PROpolypeptide variants include, for instance, PRO polypeptides wherein oneor more amino acid residues are added, or deleted, at the N- orC-terminus of the full-length native amino acid sequence. Ordinarily, aPRO polypeptide variant will have at least about 80% amino acid sequenceidentity, alternatively at least about 81% amino acid sequence identity,alternatively at least about 82% amino acid sequence identity,alternatively at least about 83% amino acid sequence identity,alternatively at least about 84% amino acid sequence identity,alternatively at least about 85% amino acid sequence identity,alternatively at least about 86% amino acid sequence identity,alternatively at least about 87% amino acid sequence identity,alternatively at least about 88% amino acid sequence identity,alternatively at least about 89% amino acid sequence identity,alternatively at least about 90% amino acid sequence identity,alternatively at least about 91% amino acid sequence identity,alternatively at least about 92% amino acid sequence identity,alternatively at least about 93% amino acid sequence identity,alternatively at least about 94% amino acid sequence identity,alternatively at least about 95% amino acid sequence identity,alternatively at least about 96% amino acid sequence identity,alternatively at least about 97% amino acid sequence identity,alternatively at least about 98% amino acid sequence identity andalternatively at least about 99% amino acid sequence identity to afull-length native sequence PRO polypeptide sequence as disclosedherein, a PRO polypeptide sequence lacking the signal peptide asdisclosed herein, an extracellular domain of a PRO polypeptide, with orwithout the signal peptide, as disclosed herein or any otherspecifically defined fragment of a full-length PRO polypeptide sequenceas disclosed herein. Ordinarily, PRO variant polypeptides are at leastabout 10 amino acids in length, alternatively at least about 20 aminoacids in length, alternatively at least about 30 amino acids in length,alternatively at least about 40 amino acids in length, alternatively atleast about 50 amino acids in length, alternatively at least about 60amino acids in length, alternatively at least about 70 amino acids inlength, alternatively at least about 80 amino acids in length,alternatively at least about 90 amino acids in length, alternatively atleast about 100 amino acids in length, alternatively at least about 150amino acids in length, alternatively at least about 200 amino acids inlength, alternatively at least about 300 amino acids in length, or more.

[0232] “Percent (%) amino acid sequence identity” with respect to thePRO polypeptide sequences identified herein is defined as the percentageof amino acid residues in a candidate sequence that are identical withthe amino acid residues in the specific PRO polypeptide sequence, afteraligning the sequences and introducing gaps, if necessary, to achievethe maximum percent sequence identity, and not considering anyconservative substitutions as part of the sequence identity. Alignmentfor purposes of determining percent amino acid sequence identity can beachieved in various ways that are within the skill in the art, forinstance, using publicly available computer software such as BLAST,BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the artcan determine appropriate parameters for measuring alignment, includingany algorithms needed to achieve maximal alignment over the full lengthof the sequences being compared. For purposes herein, however, % aminoacid sequence identity values are generated using the sequencecomparison computer program ALIGN-2, wherein the complete source codefor the ALIGN-2 program is provided in Table 1 below. The ALIGN-2sequence comparison computer program was authored by Genentech, Inc. andthe source code shown in Table 1 below has been filed with userdocumentation in the U.S. Copyright Office, Washington D.C., 20559,where it is registered under U.S. Copyright Registration No. TXU510087.The ALIGN-2 program is publicly available through Genentech, Inc., SouthSan Francisco, Calif. or may be compiled from the source code providedin Table 1 below. The ALIGN-2 program should be compiled for use on aUNIX operating system, preferably digital UNIX V4.0D. All sequencecomparison parameters are set by the ALIGN-2 program and do not vary.

[0233] In situations where ALIGN-2 is employed for amino acid sequencecomparisons, the % amino acid sequence identity of a given amino acidsequence A to, with, or against a given amino acid sequence B (which canalternatively be phrased as a given amino acid sequence A that has orcomprises a certain % amino acid sequence identity to, with, or againsta given amino acid sequence B) is calculated as follows:

100 times the fraction X/Y

[0234] where X is the number of amino acid residues scored as identicalmatches by the sequence alignment program ALIGN-2 in that program'salignment of A and B, and where Y is the total number of amino acidresidues in B. It will be appreciated that where the length of aminoacid sequence A is not equal to the length of amino acid sequence B, the% amino acid sequence identity of A to B will not equal the % amino acidsequence identity of B to A. As examples of % amino acid sequenceidentity calculations using this method, Tables 2 and 3 demonstrate howto calculate the % amino acid sequence identity of the amino acidsequence designated “Comparison Protein” to the amino acid sequencedesignated “PRO”, wherein “PRO” represents the amino acid sequence of ahypothetical PRO polypeptide of interest, “Comparison Protein”represents the amino acid sequence of a polypeptide against which the“PRO” polypeptide of interest is being compared, and “X, “Y” and “Z”each represent different hypothetical amino acid residues.

[0235] Unless specifically stated otherwise, all % amino acid sequenceidentity values used herein are obtained as described in the immediatelypreceding paragraph using the ALIGN-2 computer program. However, % aminoacid sequence identity values may also be obtained as described below byusing the WU-BLAST-2 computer program (Altschul et al., Methods inEnzymology 266:460-480 (1996)). Most of the WU-BLAST-2 search parametersare set to the default values. Those not set to default values, i.e.,the adjustable parameters, are set with the following values: overlapspan=1, overlap fraction=0.125, word threshold (T)=11, and scoringmatrix=BLOSUM62. When WU-BLAST-2 is employed, a % amino acid sequenceidentity value is determined by dividing (a) the number of matchingidentical amino acid residues between the amino acid sequence of the PROpolypeptide of interest having a sequence derived from the native PROpolypeptide and the comparison amino acid sequence of interest (i.e.,the sequence against which the PRO polypeptide of interest is beingcompared which may be a PRO variant polypeptide) as determined byWU-BLAST-2 by (b) the total number of amino acid residues of the PROpolypeptide of interest. For example, in the statement “a polypeptidecomprising an the amino acid sequence A which has or having at least 80%amino acid sequence identity to the amino acid sequence B”, the aminoacid sequence A is the comparison amino acid sequence of interest andthe amino acid sequence B is the amino acid sequence of the PROpolypeptidc of interest.

[0236] Percent amino acid sequence identity may also be determined usingthe sequence comparison program NCBI-BLAST2 (Altschul et al., NucleicAcids Res. 25:3389-3402 (1997)). The NCB1-BLAST2 sequence comparisonprogram may be downloaded from httq://www.ncbi.nlmi.nih.gov or otherwiseobtained from the National Institute of Health, Bethesda, Md.NCBI-BLAST2 uses several search parameters, wherein all of those searchparameters are set to default values including, for example, unmask=yes,strand=all, expected occurrences=10, minimum low complexity length=15/5,multi-pass e-value=0.01, constant for multi-pass =25, dropoff for finalgapped alignment=25 and scoring matrix=BLOSUM62.

[0237] In situations where NCBI-BLAST2 is employed for amino acidsequence comparisons, the % amino acid sequence identity of a givenamino acid sequence A to, with, or against a given amino acid sequence B(which can alternatively be phrased as a given amino acid sequence Athat has or comprises a certain % amino acid sequence identity to, with,or against a given amino acid sequence B) is calculated as follows:

100 times the fraction X/Y

[0238] where X is the number of amino acid residues scored as identicalmatches by the sequence alignment program NCBI-BLAST2 in that program'salignment of A and B, and where Y is the total number of amino acidresidues in B. It will be appreciated that where the length of aminoacid sequence A is not equal to the length of amino acid sequence B, the% amino acid sequence identity of A to B will not equal the % amino acidsequence identity of B to A.

[0239] “PRO variant polynucleotide” or “PRO variant nucleic acidsequence” means a nucleic acid molecule which encodes an active PROpolypeptide as defined below and which has at least about 80% nucleicacid sequence identity with a nucleotide acid sequence encoding afull-length native sequence PRO polypeptide sequence as disclosedherein, a full-length native sequence PRO polypeptide sequence lackingthe signal peptide as disclosed herein, an extracellular domain of a PROpolypeptide, with or without the signal peptide, as disclosed herein orany other fragment of a full-length PRO polypeptide sequence asdisclosed herein. Ordinarily, a PRO variant polynucleotide will have atleast about 80% nucleic acid sequence identity, alternatively at leastabout 81% nucleic acid sequence identity, alternatively at least about82% nucleic acid sequence identity, alternatively at least about 83%nucleic acid sequence identity, alternatively at least about 84% nucleicacid sequence identity, alternatively at least about 85% nucleic acidsequence identity, alternatively at least about 86% nucleic acidsequence identity, alternatively at least about 87% nucleic acidsequence identity, alternatively at least about 88% nucleic acidsequence identity, alternatively at least about 89% nucleic acidsequence identity, alternatively at least about 90% nucleic acidsequence identity, alternatively at least about 91% nucleic acidsequence identity, alternatively at least about 92% nucleic acidsequence identity, alternatively at least about 93% nucleic acidsequence identity, alternatively at least about 94% nucleic acidsequence identity, alternatively at least about 95% nucleic acidsequence identity, alternatively at least about 96% nucleic acidsequence identity, alternatively at least about 97% nucleic acidsequence identity, alternatively at least about 98% nucleic acidsequence identity and alternatively at least about 99% nucleic acidsequence identity with a nucleic acid sequence encoding a full-lengthnative sequence PRO polypeptide sequence as disclosed herein, afull-length native sequence PRO polypeptide sequence lacking the signalpeptide as disclosed herein, an extracellular domain of a PROpolypeptide, with or without the signal sequence, as disclosed herein orany other fragment of a full-length PRO polypeptide sequence asdisclosed herein. Variants do not encompass the native nucleotidesequence.

[0240] Ordinarily, PRO variant polynucleotides are at least about 30nucleotides in length, alternatively at least about 60 nucleotides inlength, alternatively at least about 90 nucleotides in length,alternatively at least about 120 nucleotides in length, alternatively atleast about 150 nucleotides in length, alternatively at least about 180nucleotides in length, alternatively at least about 210 nucleotides inlength, alternatively at least about 240 nucleotides in length,alternatively at least about 270 nucleotides in length, alternatively atleast about 300 nucleotides in length, alternatively at least about 450nucleotides in length, alternatively at least about 600 nucleotides inlength, alternatively at least about 900 nucleotides in length, or more.

[0241] “Percent (%) nucleic acid sequence identity” with respect toPRO-encoding nucleic acid sequences identified herein is defined as thepercentage of nucleotides in a candidate sequence that are identicalwith the nucleotides in the PRO nucleic acid sequence of interest, afteraligning the sequences and introducing gaps, if necessary, to achievethe maximum percent sequence identity. Alignment for purposes ofdetermining percent nucleic acid sequence identity can be achieved invarious ways that are within the skill in the art, for instance, usingpublicly available computer software such as BLAST, BLAST-2, ALIGN orMegalign (DNASTAR) software. For purposes herein, however, % nucleicacid sequence identity values are generated using the sequencecomparison computer program ALIGN-2, wherein the complete source codefor the ALIGN-2 program is provided in Table I below. The ALIGN-2sequence comparison computer program was authored by Genentech, Inc. andthe source code shown in Table I below has been filed with userdocumentation in the U.S. Copyright Office, Washington D.C., 20559,where it is registered under U.S. Copyright Registration No. TXU510087.The ALIGN-2 program is publicly available through Genentech, Inc., SouthSan Francisco, Calif. or may be compiled from the source code providedin Table 1 below. The ALIGN-2 program should be compiled for use on aUNIX operating system, preferably digital UNIX V4.0D. All sequencecomparison parameters are set by the ALIGN-2 program and do not vary.

[0242] In situations where ALIGN-2 is employed for nucleic acid sequencecomparisons, the % nucleic acid sequence identity of a given nucleicacid sequence C to, with, or against a given nucleic acid sequence D(which can alternatively be phrased as a given nucleic acid sequence Cthat has or comprises a certain % nucleic acid sequence identity to,with, or against a given nucleic acid sequence D) is calculated asfollows:

100 times the fraction W/Z

[0243] where W is the number of nucleotides scored as identical matchesby the sequence alignment program ALIGN-2 in that program's alignment ofC and D, and where Z is the total number of nucleotides in D. It will beappreciated that where the length of nucleic acid sequence C is notequal to the length of nucleic acid sequence D, the % nucleic acidsequence identity of C to D will not equal the % nucleic acid sequenceidentity of D to C. As examples of % nucleic acid sequence identitycalculations, Tables 4 and 5, demonstrate how to calculate the % nucleicacid sequence identity of the nucleic acid sequence designated“Comparison DNA” to the nucleic acid sequence designated “PRO-DNA”,wherein “PRO-DNA” represents a hypothetical PRO-encoding nucleic acidsequence of interest, “Comparison DNA” represents the nucleotidesequence of a nucleic acid molecule against which the “PRO-DNA” nucleicacid molecule of interest is being compared, and “N”, “L” and “V” eachrepresent different hypothetical nucleotides.

[0244] Unless specifically stated otherwise, all % nucleic acid sequenceidentity values used herein are obtained as described in the immediatelypreceding paragraph using the ALIGN-2 computer program. However, %nucleic acid sequence identity values may also be obtained as describedbelow by using the WU-BLAST-2 computer program (Altschul et al., Methodsin Enzymology 266:460-480 (1996)). Most of the WU-BLAST-2 searchparameters are set to the default values. Those not set to defaultvalues, i.e., the adjustable parameters, are set with the followingvalues: overlap span=1, overlap fraction=0.125, word threshold (T)=11,and scoring matrix= BLOSUM62. When WU-BLAST-2 is employed, a % nucleicacid sequence identity value is determined by dividing (a) the number ofmatching identical nucleotides between the nucleic acid sequence of thePRO polypeptide-encoding nucleic acid molecule of interest having asequence derived from the native sequence PRO polypeptide-encodingnucleic acid and the comparison nucleic acid molecule of interest (i.e.,the sequence against which the PRO polypeptide-encoding nucleic acidmolecule of interest is being compared which may be a variant PROpolynucleotide) as determined by WU-BLAST-2 by (b) the total number ofnucleotides of the PRO polypeptide-encoding nucleic acid molecule ofinterest. For example, in the statement “an isolated nucleic acidmolecule comprising a nucleic acid sequence A which has or having atleast 80% nucleic acid sequence identity to the nucleic acid sequenceB”, the nucleic acid sequence A is the comparison nucleic acid moleculeof interest and the nucleic acid sequence B is the nucleic acid sequenceof the PRO polypeptide-encoding nucleic acid molecule of interest.

[0245] Percent nucleic acid sequence identity may also be determinedusing the sequence comparison program NCBI-BLAST2 (Altschul et al.,Nucleic Acids Res. 25:3389-3402 (1997)). The NCBI-BLAST2 sequencecomparison program may be downloaded from http://www.ncbi.nlm.nih.gov orotherwise obtained from the National Institute of Health, Bethesda, Md.NCBI-BLAST2 uses several search parameters, wherein all of those searchparameters are set to default values including, for example, unmask=yes,strand=all, expected occurrences=10, minimum low complexity length=15/5,multi-pass e-value=0.01, constant for multi-pass =25, dropoff for finalgapped alignment=25 and scoring matrix= BLOSUM62.

[0246] In situations where NCBI-BLAST2 is employed for sequencecomparisons, the % nucleic acid sequence identity of a given nucleicacid sequence C to, with, or against a given nucleic acid sequence D(which can alternatively be phrased as a given nucleic acid sequence Cthat has or comprises a certain % nucleic acid sequence identity to,with, or against a given nucleic acid sequence D) is calculated asfollows:

100 times the fraction W/Z

[0247] where W is the number of nucleotides scored as identical matchesby the sequence alignment program NCBI-BLAST2 in that program'salignment of C and D, and where Z is the total number of nucleotides inD. It will be appreciated that where the length of nucleic acid sequenceC is not equal to the length of nucleic acid sequence D, the % nucleicacid sequence identity of C to D will not equal the % nucleic acidsequence identity of D to C.

[0248] In other embodiments, PRO variant polynucleotides are nucleicacid molecules that encode an active PRO polypeptide and which arecapable of hybridizing, preferably under stringent hybridization andwash conditions, to nucleotide sequences encoding a full-length PROpolypeptide as disclosed herein. PRO variant polypeptides may be thosethat are encoded by a PRO variant polynucleotide.

[0249] The term “positives”, in the context of sequence comparisonperformed as described above, includes residues in the sequencescompared that are not identical but have similar properties (e.g. as aresult of conservative substitutions, see Table 6 below). For purposesherein, the % value of positives is determined by dividing (a) thenumber of amino acid residues scoring a positive value between the PROpolypeptide amino acid sequence of interest having a sequence derivedfrom the native PRO polypeptide sequence and the comparison amino acidsequence of interest (i.e., the amino acid sequence against which thePRO polypeptide sequence is being compared) as determined in theBLOSUM62 matrix of WU-BLAST-2 by (b) the total number of amino acidresidues of the PRO polypeptide of interest.

[0250] Unless specifically stated otherwise, the % value of positives iscalculated as described in the immediately preceding paragraph. However,in the context of the amino acid sequence identity comparisons performedas described for ALIGN-2 and NCBI-BLAST-2 above, includes amino acidresidues in the sequences compared that are not only identical, but alsothose that have similar properties. Amino acid residues that score apositive value to an amino acid residue of interest are those that areeither identical to the amino acid residue of interest or are apreferred substitution (as defined in Table 6 below) of the amino acidresidue of interest.

[0251] For amino acid sequence comparisons using ALIGN-2 or NCBI-BLAST2,the % value of positives of a given amino acid sequence A to, with, oragainst a given amino acid sequence B (which can alternatively bephrased as a given amino acid sequence A that has or comprises a certain% positives to, with, or against a given amino acid sequence B) iscalculated as follows:

100 times the fraction X/Y

[0252] where X is the number of amino acid residues scoring a positivevalue as defined above by the sequence alignment program ALIGN-2 orNCBI-BLAST2 in that program's alignment of A and B, and where Y is thetotal number of amino acid residues in B. It will be appreciated thatwhere the length of amino acid sequence A is not equal to the length ofamino acid sequence B, the % positives of A to B will not equal the %positives of B to A.

[0253] “Isolated,” when used to describe the various polypeptidesdisclosed herein, means polypeptide that has been identified andseparated and/or recovered from a component of its natural environment.Contaminant components of its natural environment are materials thatwould typically interfere with diagnostic or therapeutic uses for thepolypeptide, and may include enzymes, hormones, and other proteinaceousor non-proteinaceous solutes. In preferred embodiments, the polypeptidewill be purified (1) to a degree sufficient to obtain at least 15residues of N-terminal or internal amino acid sequence by use of aspinning cup sequenator, or (2) to homogeneity by SDS-PAGE undernon-reducing or reducing conditions using Coomassie blue or, preferably,silver stain. Isolated polypeptide includes polypeptide in situ withinrecombinant cells, since at least one component of the PRO polypeptidenatural environment will not be present. Ordinarily, however, isolatedpolypeptide will be prepared by at least one purification step.

[0254] An “isolated” PRO polypeptide-encoding nucleic acid or otherpolypeptide-encoding nucleic acid is a nucleic acid molecule that isidentified and separated from at least one contaminant nucleic acidmolecule with which it is ordinarily associated in the natural source ofthe polypeptide-encoding nucleic acid. An isolated polypeptide-encodingnucleic acid molecule is other than in the form or setting in which itis found in nature. Isolated polypeptide-encoding nucleic acid moleculestherefore are distinguished from the specific polypeptide-encodingnucleic acid molecule as it exists in natural cells. However, anisolated polypeptide-encoding nucleic acid molecule includespolypeptide-encoding nucleic acid molecules contained in cells thatordinarily express the polypeptide where, for example, the nucleic acidmolecule is in a chromosomal location different from that of naturalcells.

[0255] The term “control sequences” refers to DNA sequences necessaryfor the expression of an operably linked coding sequence in a particularhost organism. The control sequences that are suitable for prokaryotes,for example, include a promoter, optionally an operator sequence, and aribosome binding site. Eukaryotic cells are known to utilize promoters,polyadenylation signals, and enhancers.

[0256] Nucleic acid is “operably linked” when it is placed into afunctional relationship with another nucleic acid sequence. For example,DNA for a presequence or secretory leader is operably linked to DNA fora polypeptide if it is expressed as a preprotein that participates inthe secretion of the polypeptide; a promoter or enhancer is operablylinked to a coding sequence if it affects the transcription of thesequence; or a ribosome binding site is operably linked to a codingsequence if it is positioned so as to facilitate translation. Generally,“operably linked” means that the DNA sequences being linked arecontiguous, and, in the case of a secretory leader, contiguous and inreading phase. However, enhancers do not have to be contiguous. Linkingis accomplished by ligation at convenient restriction sites. If suchsites do not exist, the synthetic oligonucleotide adaptors or linkersare used in accordance with conventional practice.

[0257] The term “antibody” is used in the broadest sense andspecifically covers, for example, single anti-PRO monoclonal antibodies(including agonist, antagonist, and neutralizing antibodies), anti-PROantibody compositions with polyepitopic specificity, single chainanti-PRO antibodies, and fragments of anti-PRO antibodies (see below).The term “monoclonal antibody” as used herein refers to an antibodyobtained from a population of substantially homogeneous antibodies,i.e., the individual antibodies comprising the population are identicalexcept for possible naturally-occurring mutations that may be present inminor amounts.

[0258] “Stringency” of hybridization reactions is readily determinableby one of ordinary skill in the art, and generally is an empiricalcalculation dependent upon probe length, washing temperature, and saltconcentration. In general, longer probes require higher temperatures forproper annealing, while shorter probes need lower temperatures.Hybridization generally depends on the ability of denatured DNA toreanneal when complementary strands are present in an environment belowtheir melting temperature. The higher the degree of desired homologybetween the probe and hybridizable sequence, the higher the relativetemperature which can be used. As a result, it follows that higherrelative temperatures would tend to make the reaction conditions morestringent, while lower temperatures less so. For additional details andexplanation of stringency of hybridization reactions, see Ausubel etal., Current Protocols in Molecular Biology, Wiley lntersciencePublishers, (1995).

[0259] “Stringent conditions” or “high stringency conditions”, asdefined herein, may be identified by those that: (1) employ low ionicstrength and high temperature for washing, for example 0.015 M sodiumchloride/0.0015 M sodium citrate/0.1% sodium dodecyl sulfate at 50° C.;(2) employ during hybridization a denaturing agent, such as formamide,for example, 50% (v/v) formamide with 0.1% bovine serum albumin/0.1%Ficoll/0.1% polyvinylpyrrolidone/50 mM sodium phosphate buffer at pH 6.5with 750 mM sodium chloride, 75 mM sodium citrate at 42° C.; or (3)employ 50% formamide, 5×SSC (0.75 M NaCl, 0.075 M sodium citrate), 50 mMsodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5× Denhardt'ssolution, sonicated salmon sperm DNA (50 μg/ml), 0.1% SDS, and 10%dextran sulfate at 42° C., with washes at 42° C. in 0.2×SSC (sodiumchloride/sodium citrate) and 50% formamide at 55° C., followed by ahigh-stringency wash consisting of 0.1×SSC containing EDTA at 55° C.

[0260] “Moderately stringent conditions” may be identified as describedby Sambrook et al., Molecular Cloning: A Laboratory Manual, New York:Cold Spring Harbor Press, 1989, and include the use of washing solutionand hybridization conditions (e.g., temperature, ionic strength and %SDS) less stringent that those described above. An example of moderatelystringent conditions is overnight incubation at 37° C. in a solutioncomprising: 20% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate),50 mM sodium phosphate (pH 7.6), 5× Denhardt's solution, 10% dextransulfate, and 20 mg/ml denatured sheared salmon sperm DNA, followed bywashing the filters in 1×SSC at about 37-50° C. The skilled artisan willrecognize how to adjust the temperature, ionic strength, etc. asnecessary to accommodate factors such as probe length and the like.

[0261] The term “epitope tagged” when used herein refers to a chimericpolypeptide comprising a PRO polypeptide fused to a “tag polypeptide”.The tag polypeptide has enough residues to provide an epitope againstwhich an antibody can be made, yet is short enough such that it does notinterfere with activity of the polypeptide to which it is fused. The tagpolypeptide preferably also is fairly unique so that the antibody doesnot substantially cross-react with other epitopes. Suitable tagpolypeptides generally have at least six amino acid residues and usuallybetween about 8 and 50 amino acid residues (preferably, between about 10and 20 amino acid residues).

[0262] As used herein, the term “immunoadhesin” designates antibody-likemolecules which combine the binding specificity of a heterologousprotein (an “adhesin”) with the effector functions of immunoglobulinconstant domains. Structurally, the immunoadhesins comprise a fusion ofan amino acid sequence with the desired binding specificity which isother than the antigen recognition and binding site of an antibody(i.e., is “heterologous”), and an immunoglobulin constant domainsequence. The adhesin part of an immunoadhesin molecule typically is acontiguous amino acid sequence comprising at least the binding site of areceptor or a ligand. The immunoglobulin constant domain sequence in theimmunoadhesin may be obtained from any immunoglobulin, such as IgG-1,IgG-2, IgG-3, or IgG-4 subtypes, IgA (including IgA-1 and IgA-2), IgE,IgD or IgM.

[0263] “Active” or “activity” for the purposes herein refers to form(s)of a PRO polypeptide which retain a biological and/or an immunologicalactivity of native or naturally-occurring PRO, wherein “biological”activity refers to a biological function (either inhibitory orstimulatory) caused by a native or naturally-occurring PRO other thanthe ability to induce the production of an antibody against an antigenicepitope possessed by a native or naturally-occurring PRO and an“immunological” activity refers to the ability to induce the productionof an antibody against an antigenic epitope possessed by a native ornaturally-occurring PRO.

[0264] The term “antagonist” is used in the broadest sense, and includesany molecule that partially or fully blocks, inhibits, or neutralizes abiological activity of a native PRO polypeptide disclosed herein. In asimilar manner, the term “agonist” is used in the broadest sense andincludes any molecule that mimics a biological activity of a native PROpolypeptide disclosed herein. Suitable agonist or antagonist moleculesspecifically include agonist or antagonist antibodies or antibodyfragments, fragments or amino acid sequence variants of native PROpolypeptides, peptides, antisense oligonucleotides, small organicmolecules, etc. Methods for identifying agonists or antagonists of a PROpolypeptide may comprise contacting a PRO polypeptide with a candidateagonist or antagonist molecule and measuring a detectable change in oneor more biological activities normally associated with the PROpolypeptide.

[0265] “Treatment” refers to both therapeutic treatment and prophylacticor preventative measures, wherein the object is to prevent or slow down(lessen) the targeted pathologic condition or disorder. Those in need oftreatment include those already with the disorder as well as those proneto have the disorder or those in whom the disorder is to be prevented.

[0266] “Chronic” administration refers to administration of the agent(s)in a continuous mode as opposed to an acute mode, so as to maintain theinitial therapeutic effect (activity) for an extended period of time.“Intermittent” administration is treatment that is not consecutivelydone without interruption, but rather is cyclic in nature.

[0267] “Mammal” for purposes of treatment refers to any animalclassified as a mammal, including humans, domestic and farm animals, andzoo, sports, or pet animals, such as dogs, cats, cattle, horses, sheep,pigs, goats, rabbits, etc. Preferably, the mammal is human.

[0268] Administration “in combination with” one or more furthertherapeutic agents includes simultaneous (concurrent) and consecutiveadministration in any order.

[0269] “Carriers” as used herein include pharmaceutically acceptablecarriers, excipients, or stabilizers which are nontoxic to the cell ormammal being exposed thereto at the dosages and concentrations employed.Often the physiologically acceptable carrier is an aqueous pH bufferedsolution. Examples of physiologically acceptable carriers includebuffers such as phosphate, citrate, and other organic acids;antioxidants including ascorbic acid; low molecular weight (less thanabout 10 residues) polypeptide; proteins, such as serum albumin,gelatin, or immunoglobulin; hydrophilic polymers such aspolyvinylpyrrolidone; amino acids such as glycine, glutamine,asparagine, arginine or lysine; monosaccharides, disaccharides, andother carbohydrates including glucose, mannose, or dextrins; chelatingagents such as EDTA; sugar alcohols such as mannitol or sorbitol;salt-forming counterions such as sodium; and/or nonionic surfactantssuch as TWEEN™, polyethylene glycol (PEG), and PLURONICS™.

[0270] “Antibody fragments” comprise a portion of an intact antibody,preferably the antigen binding or variable region of the intactantibody. Examples of antibody fragments include Fab, Fab′, F(ab′)₂, andFv fragments; diabodies; linear antibodies (Zapata et al., Protein Eng.8(10): 1057-1062 [1995]); single-chain antibody molecules; andmultispecific antibodies formed from antibody fragments.

[0271] Papain digestion of antibodies produces two identicalantigen-binding fragments, called “Fab” fragments, each with a singleantigen-binding site, and a residual “Fc” fragment, a designationreflecting the ability to crystallize readily. Pepsin treatment yieldsan F(ab′)₂ fragment that has two antigen-combining sites and is stillcapable of cross-linking antigen.

[0272] “Fv” is the minimum antibody fragment which contains a completeantigen-recognition and -binding site. This region consists of a dimerof one heavy- and one light-chain variable domain in tight, non-covalentassociation. It is in this configuration that the three CDRs of eachvariable domain interact to define an antigen-binding site on thesurface of the V_(H)-V_(L) dimer. Collectively, the six CDRs conferantigen-binding specificity to the antibody. However, even a singlevariable domain (or half of an Fv comprising only three CDRs specificfor an antigen) has the ability to recognize and bind antigen, althoughat a lower affinity than the entire binding site.

[0273] The Fab fragment also contains the constant domain of the lightchain and the first constant domain (CH1) of the heavy chain. Fabfragments differ from Fab′ fragments by the addition of a few residuesat the carboxy terminus of the heavy chain CH1 domain including one ormore cysteines from the antibody hinge region. Fab′—SH is thedesignation herein for Fab′ in which the cysteine residue(s) of theconstant domains bear a free thiol group. F(ab′)₂ antibody fragmentsoriginally were produced as pairs of Fab′ fragments which have hingecysteines between them. Other chemical couplings of antibody fragmentsare also known.

[0274] The “light chains” of antibodies (immunoglobulin) from anyvertebrate species can be assigned to one of two clearly distinct types,called kappa and lambda, based on the amino acid sequences of theirconstant domains.

[0275] Depending on the amino acid sequence of the constant domain oftheir heavy chains, immunoglobulin can be assigned to different classes.There are five major classes of immunoglobulin: IgA, IgD, IgE, IgG, andIgM, and several of these may be further divided into subclasses(isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA, and IgA2.

[0276] “Single-chain Fv” or “sFv” antibody fragments comprise the V_(H)and V_(L) domains of antibody, wherein these domains are present in asingle polypeptide chain. Preferably, the Fv polypeptide furthercomprises a polypeptide linker between the V_(H) and V_(L) domains whichenables the sFv to form the desired structure for antigen binding. For areview of sFv, see Pluckthun in The Pharmacology of MonoclonalAntibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, NewYork, pp. 269-315 (1994).

[0277] The term “diabodies” refers to small antibody fragments with twoantigen-binding sites, which fragments comprise a heavy-chain variabledomain (V_(H)) connected to a light-chain variable domain (V_(L)) in thesame polypeptide chain (V_(H)-V_(L)). By using a linker that is tooshort to allow pairing between the two domains on the same chain, thedomains are forced to pair with the complementary domains of anotherchain and create two antigen-binding sites. Diabodies are described morefully in, for example, EP 404,097; WO 93/11161; and Hollinger et al.,Proc. NatI. Acad. Sci. USA, 90:6444-6448 (1993).

[0278] An “isolated” antibody is one which has been identified andseparated and/or recovered from a component of its natural environment.Contaminant components of its natural environment are materials whichwould interfere with diagnostic or therapeutic uses for the antibody,and may include enzymes, hormones, and other proteinaceous ornonproteinaceous solutes. In preferred embodiments, the antibody will bepurified (1) to greater than 95% by weight of antibody as determined bythe Lowry method, and most preferably more than 99% by weight, (2) to adegree sufficient to obtain at least 15 residues of N-terminal orinternal amino acid sequence by use of a spinning cup sequenator, or (3)to homogeneity by SDS-PAGE under reducing or nonreducing conditionsusing Coomassie blue or, preferably, silver stain. Isolated antibodyincludes the antibody in situ within recombinant cells since at leastone component of the antibody's natural environment will not be present.Ordinarily, however, isolated antibody will be prepared by at least onepurification step.

[0279] The word “label” when used herein refers to a detectable compoundor composition which is conjugated directly or indirectly to theantibody so as to generate a “labeled” antibody. The label may bedetectable by itself (e.g. radioisotope labels or fluorescent labels)or, in the case of an enzymatic label, may catalyze chemical alterationof a substrate compound or composition which is detectable.

[0280] By “solid phase” is meant a non-aqueous matrix to which theantibody of the present invention can adhere. Examples of solid phasesencompassed herein include those formed partially or entirely of glass(e.g., controlled pore glass), polysaecharides (e.g., agarose),polyacrylamides, polystyrene, polyvinyl alcohol and silicones. Incertain embodiments, depending on the context, the solid phase cancomprise the well of an assay plate; in others it is a purificationcolumn (e.g., an affinity chromatography column). This term alsoincludes a discontinuous solid phase of discrete particles, such asthose described in U.S. Pat. No. 4,275,149.

[0281] A “liposome” is a small vesicle composed of various types oflipids, phospholipids and/or surfactant which is useful for delivery ofa drug (such as a PRO polypeptide or antibody thereto) to a mammal. Thecomponents of the liposome are commonly arranged in a bilayer formation,similar to the lipid arrangement of biological membranes.

[0282] A “small molecule” is defined herein to have a molecular weightbelow about 500 Daltons.

[0283] “FGFR-1”, “FGFR-2”, “FGFR-3” and FGFR-4” refer to the fibroblastgrowth factor receptors 1, 2, 3 and 4, respectively, as disclosed byIsacchi et al., Nuc. Acids Res. 18(7):1906 (1990), Dionne et al., EMBOJ. 9(9):2685-2692 (1990), Keegan et al., Proc. Natl. Acad. Sci. USA88:1095-1099 (1991) an Partanen et al., EMBO J. 10(6):1347-1354 (1991),respectively. TABLE 1 /*  *  * C-C increased from 12 to 15  * Z isaverage of EQ  * B is average of ND  * match with stop is _M; stop-stop= 0; J (joker) match = 0  */ #define _M −8 /* value of a match with astop */ int _day[26][26] = { /*  A B C D E F G H I J K L M N O P Q R S TU V W X Y Z */ /* A */ {2, 0, −2, 0, 0, −4, 1, −1, −1, 0, −1, −2, −1, 0,_M, 1, 0, −2, 1, 1, 0, 0, −6, 0, −3, 0}, /* B */ {0, 3, −4, 3, 2, −5, 0,1, −2, 0, 0, −3, −2, 2, _M, −1, 1, 0, 0, 0, 0, −2, −5, 0, −3, 1}, /* C*/ {−2, −4, 15, −5, −5, −4, −3, −3, −2, 0, −5, −6, −5, −4, _M, −3, −5,−4, 0, −2, 0, −2, −8, 0, 0, −5}, /* D */ {0, 3, −5, 4, 3, −6, 1, 1, −2,0, 0, −4, −3, 2, _M, −1, 2, −1, 0, 0, 0, −2, −7, 0, −4, 2}, /* E */ {0,2, −5, 3, 4, −5, 0, 1, −2, 0, 0, −3, −2, 1, _M, −1, 2, −1, 0, 0, 0, −2,−7, 0, −4, 3}, /* F */ {−4, −5, −4, −6, −5, 9, −5, −2, 1, 0, −5, 2, 0,−4, _M, −5, −5, −4, −3, −3, 0, −1, 0, 0, 7, −5}, /* G */ {1, 0, −3, 1,0, −5, 5, −2, −3, 0, −2, −4, −3, 0, _M, −1, −1, −3, 1, 0, 0, −1, −7, 0,−5, 0}, /* H */ {−1, 1, −3, 1, 1, −2, −2, 6, −2, 0, 0, −2, −2, 2, _M, 0,3, 2, −1, −1, 0, −2, −3, 0, 0, 2}, /* I */ {−1, −2, −2, −2, −2, 1, −3,−2, 5, 0, −2, 2, 2, −2, _M, −2, −2, −2, −1, 0, 0, 4, −5, 0, −1, −2}, /*J */ {0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, _M, 0, 0, 0, 0, 0, 0, 0,0, 0, 0, 0}, /* K */ {−1, 0, −5, 0, 0, −5, −2, 0, −2, 0, 5, −3, 0, 1,_M, −1, 1, 3, 0, 0, 0, −2, −3, 0, −4, 0}, /* L */ {−2, −3, −6, −4, −3,2, −4, −2, 2, 0, −3, 6, 4, −3, _M, −3, −2, −3, −3 , −1, 0, 2, −2, 0, −1,−2} /* M */ {−1, −2, −5, −3, −2, 0, −3, −2, 2, 0, 0, 4, 6, −2, _M, −2,−1, 0, −2, −1, 0, 2, −4, 0, −2, −1}, /* N */ {0, 2, −4, 2, 1, −4, 0, 2,−2, 0, 1, −3, −2, 2, _M, −1, 1, 0, 1, 0, 0, −2, −4, 0, −2, 1}, /* O */{_M,_M,_M,_M,_M,_M,_M,_M,_M,_M,_M,_M,_M,_M,0,_M,_M,_M,_M,_M,_M,_M,_M,_M,_M,_M,}, /* P */ {1, −1, −3, −1, −1, −5,−1, 0, −2, 0, −1, −3, −2, −1,_M, 6, 0, 0, 1, 0, 0, −1, −6, 0, −5, 0}, /*Q */ {0, 1, −5, 2, 2, −5, −1, 3, −2, 0, 1, −2, −1, 1, _M, 0, 4, 1, −1,−1, 0, −2, −5, 0, −4, 3}, /* R */ {−2, 0, −4, −1, −1, −4, −3, 2, −2, 0,3, −3, 0, 0, _M, 0, 1, 6, 0, −1, 0, −2, 2, 0, −4, 0}, /* S */ {1, 0, 0,0, 0, −3, 1, −1, −1, 0, 0, −3, −2, 1, _M, 1, −1, 0, 2, 1, 0, −1, −2, 0,−3, 0}, /* T */ {1, 0, −2, 0, 0, −3, 0, −1, 0, 0, 0, −1, −1, 0, _M, 0,−1, −1, 1, 3, 0, 0, −5, 0, −3, 0}, /* U */ {0, 0, 0, 0, 0, 0, 0, 0, 0,0, 0, 0, 0, 0,_M, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0}, /* V */ {0, −2, −2,−2, −2, −1, −1, −2, 4, 0, −2, 2, 2, −2,_M, −1, −2, −2, −1, 0, 0, 4, −6,0, −2, −2}, /* W */ {−6, −5, −8, −7, −7, 0, −7, −3, −5, 0, −3, −2, −4,−4,_M, −6, −5, 2, −2, −5, 0, −6, 17, 0, 0, −6}, /* X */ {0, 0, 0, 0, 0,0, 0, 0, 0, 0, 0, 0, 0, 0, _M, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0}, /* Y */{−3, −3, 0, −4, −4, 7, −5, 0, −1, 0, −4, −1, −2, −2, _M, −5, −4, −4, −3,−3, 0, −2, 0, 0, 10, −4}, /* Z */ {0, 1, −5, 2, 3, −5, 0, 2, −2, 0, 0,−2, −1, 1,_M, 0, 3, 0, 0, 0, 0, −2, −6, 0, −4, 4}, }; /*  */ #include<stdio.h> #include <ctype.h> #define MAXJMP  16 /* max jumps in a diag*/ #define MAXGAP  24 /* don't continue to penalize gaps larger thanthis */ #define JMPS 1024 /* max jmps in an path */ #define MX   4 /*save if there's at least MX-1 bases since last jmp */ #define DMAT   3/* value of matching bases */ #define DMIS   0 /* penalty for mismatchedbases */ #define DINS0   8 /* penalty for a gap */ #define DINS1   1 /*penalty per base */ #define PINS0   8 /* penalty for a gap */ #definePINS1   4 /* penalty per residue */ struct jmp { short n[MAXJMP]; /*size of jmp (neg for dely) */ unsigned short x[MAXJMP]; /* base no. ofjmp in seq x */ /* limits seq to 2{circumflex over ( )}16 −1 */ };struct diag { int score; /* score at last jmp */ long offset; /* offsetof prev block */ short ijmp; /* current jmp index */ struct jmp jp; /*list of jmps */ }; struct path { int spc; /* number of leading spaces */short n[JMPS]; /* size of jmp (gap) */ int x[JMPS]; /* loc of jmp (lastelem before gap) */ }; char *ofile; /* output file name */ char*namex[2]; /* seq names: getseqs() */ char *prog; /* prog name for errmsgs */ char *seqx[2];   /* seqs: getseqs() */ int dmax; /* best diag:nw() */ int dmax0; /* final diag */ int dna; /* set if dna: main() */int endgaps; /* set if penalizing end gaps */ int gapx, gapy; /* totalgaps in seqs */ int len0, len1; /* seq lens */ int ngapx, ngapy; /*total size of gaps */ int smax; /* max score: nw() */ int *xbm; /*bitmap for matching */ long offset; /* current offset in jmp file */struct diag *dx; /* holds diagonals */ struct path pp[2]; /* holds pathfor seqs */ char *calloc(), *malloc(), *index(), *strcpy(); char*getseq(), *g_calloc(); /* Needleman-Wunsch alignment program  *  *usage: progs file1 file2  * where file1 and file2 are two dna or twoprotein sequences.  * The sequences can be in upper- or lower-case anmay contain ambiguity  * Any lines beginning with ‘;’, ‘>’ or ‘<’ areignored  * Max file length is 65535 (limited by unsigned short x in thejmp struct)  * A sequence with ⅓ or more of its elements ACGTU isassumed to be DNA  * Output is in the file “align.out”  *  * The programmay create a tmp file in /tmp to hold info about traceback.  * Originalversion developed under BSD 4.3 on a vax 8650  */ #include “nw.h”#include “day.h” static _dbval[26] = {1,14,2,13,0,0,4,11,0,0,12,0,3,15,0,0,0,5,6,8,8,7,9,0,10,0 }; static_pbval[26] = { 1, 2|(1< <(‘D’-‘A’))|(1< <(‘N’-‘A’)), 4, 8, 16, 32, 64,128, 256, 0×FFFFFFF, 1< <10, 1< <11, 1< <12, 1< <13, 1< <14, 1< <15, 1<<16, 1< <17, 1< <18, 1< <19, 1< <20, 1< <21, 1< <22, 1< <23, 1< <24, 1<<25|(1< <(‘E’-‘A’))|(1< <(‘Q’-‘A’)) }; main(ac, av) main int ac; char*av[]; { prog = av[0]; if(ac != 3) { fprintf(stderr, “usage: %s file1file2\n”, prog); fprintf(stderr, “where file1 and file2 are two dna ortwo protein sequences.\n”); fprintf(stderr, “The sequences can be inupper- or lower-case\n”); fprintf(stderr, “Any lines beginning with ‘;’or ‘<’ are ignored\n”); fprintf(stderr, “Output is in the file\“align.out\”\n”); exit(1); } namex[0] = av[1]; namex[1] = av[2];seqx[0] = getseq(namex[0], &len0); seqx[1] = getseq(namex[1], &len1);xbm = (dna)? _dbval : _pbval; endgaps = 0; /* 1 to penalize endgaps */ofile = “align.out”; /* output file */ nw(); /* fill in the matrix, getthe possible jmps */ readjmps(); /* get the actual jmps */ print(); /*print stats, alignment */ cleanup(0); /* unlink any tmp files */ } /* dothe alignment, return best score: main()  * dna: values in Fitch andSmith, PNAS, 80, 1382-1386, 1983  * pro: PAM 250 values  * When scoresare equal, we prefer mismatches to any gap, prefer  * a new gap toextending an ongoing gap, and prefer a gap in seqx  * to a gap in seq y. */ nw() nw { char *px, *py;   /* seqs and ptrs */ int *ndely, *dely; /*keep track of dely */ int ndelx, delx; /* keep track of delx */ int*tmp; /* for swapping row0, row1 */ int mis; /* score for each type */int ins0, ins1; /* insertion penalties */ register id; /* diagonal index*/ register ij; /* jmp index */ register *col0, *col1; /* score forcurr, last row */ register xx, yy; /* index into seqs */ dx = (structdiag *)g_calloc(“to get diags”, len0+len1+1, sizeof(struct diag)); ndely= (int *)g_calloc(“to get ndely”, len1+1, sizeof(int)); dely = (int*)g_calloc(“to get dely”, len1+1, sizeof(int)); col0 = (int*)g_calloc(“to get col0”, len1+1, sizeof(int)); col1 = (int*)g_calloc(“to get col1”, len1+1, sizeof(int)); ins0 = (dna)? DINS0 :PINS0; ins1 = (dna)? DINS1 : PlNS1; smax = −10000; if (endgaps) { for(col0[0] = dely[0] = −ins0, yy = 1; yy <= len1; yy++) { col0[yy] =dely[yy] = col0[yy−1] − ins1; ndely[yy] = yy; } col0[0] = 0; /* WatermanBull Math Biol 84 */ } else for (yy= 1; yy <= len1; yy++) dely[yy] =−ins0; /* fill in match matrix  */ for (px = seqx[0], xx = 1; xx <=len0; px++, xx++) { /* initialize first entry in col  */ if (endgaps) {if (xx == 1) col1[0] = delx = −(ins0+ins1); else col1[0] = delx =col0[0]−ins1; ndelx = xx; } else { col1[0] = 0; delx = −ins0; ndelx = 0;} ...nw for (py = seqx[1], yy = 1; yy <= len1; py++, yy++) { mis =col0[yy−1]; if (dna) mis + = (xbm[*px−‘A’]&xbm[*py−‘A’])? DMAT : DMIS;else mis += _day[*px−‘A’][*py−‘A’]; /* update penalty for del in x seq; * favor new del over ongong del  * ignore MAXGAP if weighting endgaps */ if (endgaps || ndely[yy] < MAXGAP) { if (col0[yy] − ins0 >=dely[yy]) { dely[yy] = col0[yy] − (ins0+ins1); ndely[yy] = 1; } else {dely[yy] −= ins1; ndely[yy]++; } } else { if (col0[yy] − (ins0+ins1) >=dely[yy]) { dely[yy] = col0[yy] − (ins0+ins1); ndely[yy] = 1; } elsendely[yy]++; } /* update penalty for del in y seq;  * favor new del overongong del  */ if (endgaps || ndelx < MAXGAP) { if(col1[yy−1] − ins0 >=delx) { delx = col1[yy−1] − (ins0+ins1); ndelx = 1; } else { delx −=ins1; ndelx++; } } else { if (col1[yy−1] − (ins0+ins1) >= delx) { delx =col1[yy−1] − (ins0+ins1); ndelx = 1; } else ndelx++; } /* pick themaximum score; we're favoring  * mis over any del and delx over dely  */...nw id = xx − yy + len1 − 1; if (mis >= delx && mis >= dely[yy])col1[yy] = mis; else if (delx >= dely[yy]) { col1[yy] = delx; ij =dx[id].ijmp; if (dx[id].jp.n[0] && (!dna || (ndelx >= MAXJMP && xx >dx[id].jp.x[ij]+MX) || mis > dx[id].score+DINS0)) { dx[id].ijmp++; if(++ij >= MAXJMP) { writejmps(id); ij = dx[id].ijmp = 0; dx[id].offset =offset; offset += sizeof(struct jmp) + sizeof(offset); } }dx[id].jp.n[ij] = ndelx; dx[id].jp.x[ij] = xx; dx[id].score = delx; }else { col1[yy] = dely[yy]; ij = dx[id].ijmp; if (dx[id].jp.n[0] &&(!dna || (ndely[yy] >= MAXJMP && xx > dx[id].jp.x[ij]+MX) || mis >dx[id].score+DINS0)) { dx[id].ijmp++; if (++ij >= MAXJMP) {writejmps(id); ij = dx[id].ijmp = 0; dx[id].offset = offset; offset +=sizeof(struct jmp) + sizeof(offset); } } dx[id].jp.n[ij] =−ndely[yy];dx[id].jp.x[ij] = xx; dx[id].score = dely[yy]; } if (xx == len0 && yy <len1) { /* last col  */ if (endgaps) col1[yy] −= ins0+ins1*(len1−yy);if(col1[yy] > smax) { smax = col1[yy]; dmax = id; } } } if (endgaps &&xx < len0) col1[yy−1] −= ins0+ins1*(len0−xx); if (col1[yy−1] > smax) {smax = col1[yy−1]; dmax = id; } tmp = col0; col0 = col1; col1 = tmp; }(void) free((char *)ndely); (void) free((char *)dely); (void) free((char*)col0); (void) free((char *)col1); } /*  *  * print() -- only routinevisible outside this module  *  * static:  * getmat() -- trace back bestpath, count matches: print()  * pr_align() -- print alignment ofdescribed in array p[]: print()  * dumpblock() -- dump a block of lineswith numbers, stars: pr_align()  * nums() -- put out a number line:dumpblock()  * putline() -- put out a line (name, [num], seq, [num]):dumpblock()  * stars() - -put a line of stars: dumpblock()  *stripname() -- strip any path and prefix from a seqname  */ #include“nw.h” #define SPC  3 #define P_LINE 256 /* maximum output line */#define P_SPC  3 /* space between name or num and seq */ extern_day[26][26]; int olen; /* set output line length */ FILE *fx; /* outputfile */ print() print { int lx, ly, firstgap, lastgap;  /* overlap */ if((fx = fopen(ofile, “w”)) == 0) { fprintf(stderr, “%s: can't write%s\n”, prog, ofile); cleanup(1); } fprintf(fx, “<first sequence: %s(length = %d)\n”, namex[0], len0); fprintf(fx, “<second sequence: %s(length = %d)\n”, namex[1], len1); olen = 60; lx = len0; ly = len1;firstgap = lastgap = 0; if (dmax < len1 − 1) { /* leading gap in x */pp[0].spc = firstgap = len1 − dmax − 1; ly −= pp[0].spc; } else if(dmax > len1 − 1) { /* leading gap in y */ pp[1].spc = firstgap = dmax −(len1 − 1); lx −= pp[1].spc; } if (dmax0 < len0 − 1) { /* trailing gapin x */ lastgap = len0 − dmax0 −1; lx −= lastgap; } else if (dmax0 >len0 − 1) { /* trailing gap in y */ lastgap = dmax0 − (len0 − 1); ly −=lastgap; } getmat(lx, ly, firstgap, lastgap); pr_align(); } /*  * traceback the best path, count matches  */ static getmat(lx, ly, firstgap,lastgap) getmat int lx, ly; /* “core” (minus endgaps) */ int firstgap,lastgap; /* leading trailing overlap */ { int nm, i0, i1, siz0, siz1;char outx[32]; double pct; register n0, n1; register char *p0, *p1; /*get total matches, score  */ i0 = i1 = siz0 = siz1 = 0; p0 = seqx[0] +pp[1].spc; p1 = seqx[1] + pp[0].spc; n0 = pp[1].spc + 1; n1 =pp[0].spc + 1; nm = 0; while ( *p0 && *p1 ) { if (siz0) { p1++; n1++;siz0−−; } else if (siz1) { p0++; n0++; siz1−−; } else { if(xbm[*p0−‘A’]&xbm[*p1−‘A’]) nm++; if (n0++ == pp[0].x[i0]) siz0 =pp[0].n[i0++]; if (nl++ == pp[1].x[i1]) siz1 = pp[1].n[il++]; p0++;p1++; } } /* pct homology:  * if penalizing endgaps, base is the shorterseq  * else, knock off overhangs and take shorter core  */ if (endgaps)lx = (len0 < len1)? len0 : len1; else lx = (lx < ly)? lx : ly; pct =100.*(double)nm/(double)lx; fprintf(fx, “\n”); fprintf(fx, “<%d match%sin an overlap of %d: %.2f percent similarity\n”, nm, (nm == 1)? “” :“es”, lx, pct); fprintf(fx, “<gaps in first sequence: %d”, gapx);...getmat if (gapx) { (void) sprintf(outx, “(%d %s%s)”, ngapx, (dna)?“base”: “residue”, (ngapx == 1)? “”:“s”); fprintf(fx, “% s”, outx);fprintf(fx, “, gaps in second sequence: %d”, gapy); if (gapy) { (void)sprintf(outx, “(%d %s%s)”, ngapy, (dna)? “base”:“residue”, (ngapy == 1)?“”:“s”); fprintf(fx, “%s”, outx); } if (dna) fprintf(fx, “\n<score: %d(match = %d, mismatch = %d, gap penalty = %d + %d per base)\n”, smax,DMAT, DMIS, DINS0, DINS1); else fprintf(fx, “\n<score: %d (Dayhoff PAM250 matrix, gap penalty = %d + %d per residue)\n”, smax, PINS0, PINS1);if (endgaps) fprintf(fx, “<endgaps penalized. left endgap: %d %s%s,right endgap: %d %s%s\n”, firstgap, (dna)? “base” : “residue”, (firstgap== 1)? “” : “s”, lastgap, (dna)? “base” : “residue”, (lastgap == 1)? “”: “s”); else fprintf(fx, “<endgaps not penalized\n”); } static nm; /*matches in core -- for checking */ static lmax; /* lengths of strippedfile names */ static ij[2]; /* jmp index for a path */ static nc[2]; /*number at start of current line */ static ni[2]; /* current elem number-- for gapping */ static siz[2]; static char *ps[2]; /* ptr to currentelement */ static char *po[2]; /* ptr to next output char slot */ staticchar out[2][P_LINE]; /* output line */ static char star[P_LINE]; /* setby stars() */ /*  * print alignment of described in struct path pp[]  */static pr_align() pr_align { int nn; /* char count */ int more; registeri; for (i = 0, lmax = 0; i < 2++) { nn = stripname(namex[i]); if (nn >lmax) lmax = nn; nc[i] = 1; ni[i] = 1; siz[i] = ij[i] = 0; ps[i] =seqx[i]; po[i] = out[i]; } for (nn = nm = 0, more = 1; more;) {...pr_align for (i = more = 0; i < 2; i++) { /*  * do we have more ofthis sequence?  */ if (!*ps[i]) continue; more++; if (pp[i].spc) { /*leading space */ *po[i]++ = ‘ ’; pp[i].spc−−; } else if (siz[i]) { /* ina gap */ *po[i]++ = ‘−’; siz[i]−−; } else { /* we're putting a seqelement */ *po[i] = *ps[i]; if (islower(*ps[i])) *ps[i] =toupper(*ps[i]); po[i]++; ps[i]++; /*  * are we at next gap for thisseq?  */ if (ni[i] == pp[i].x[ij[i]]) { /*  * we need to merge all gaps * at this location  */ siz[i] == pp[i].n[ij[i]++]; while (ni[i] ==pp[i].x[ij[i]]) siz[i] += pp[i].n[ij[i]++]; } ni[i]++; } } if (++nn ==olen || !more && nn) { dumpblock(); for (i = 0; i < 2; i++) po[i] =out[i]; nn = 0; } } } /*  * dump a block of lines, including numbers,stars: pr_align()  */ static dumpblock() dumpblock { register i; for(i =0; i < 2; i++) *po[i]−− = ‘\0’; ...dumpblock (void) putc(‘\n’, fx); for(i = 0; i < 2; i++) { if (*out[i] && (*out[i] != ‘ ’ || *(po[i]) != ‘’)) { if (i == 0) nums(i); if (i == 0 && *out[1]) stars(); putline(i);if (i == 0 && *out[1]) fprintf(fx, star); if (i == 1) nums(i); } } } *put out a number line: dumpblock()  */ static nums(ix) nums int  ix; /*index in out[] holding seq line */ { char nline[P_LINE]; register i, j;register char *pn, *px, *py; for(pn = nline, i = 0; i < lmax+P_SPC; i++,pn++) *pn = ‘ ’; for (i = nc[ix], py = out[ix]; *py; py++, pn++) { if(*py == ‘ ’ || *py == ‘−’); *pn = ‘ ’; else { if (i%10 == 0 || (i == 1&& nc[ix] != 1)) { j = (i < 0)? −i : i; for (px = pn; j; j/= 10, px−−)*px = j%10 + ‘0’; if (i < 0) *px = ‘−’; } else *pn = ‘ ’; i++; } } *pn =‘\0’; nc[ix] = i; for (pn = nline; *pn; pn++) (void) putc(*pn, fx);(void) putc(‘\n’, fx); } /*  * put out a line (name, [num], seq. [num]):dumpblock()  */ static putline(ix) putline int   ix; { ...putline int i;register char *px; for (px = namex[ix], i = 0; *px && *px != ‘:’; px++,i++) (void) putc(*px, fx); for (;i < lmax+P_SPC; i++) (void) putc(‘ ’,fx); /* these count from 1:  * ni[] is current element (from 1)  * nc[]is number at start of current line  */ for (px = out[ix]; *px; px++)(void) putc(*px&0x7F, fx); (void) putc(‘\n’, fx); } /*  * put a line ofstars (seqs always in out[0], out[1]): dumpblock()  */ static stars()stars { int i; register char *p0, *p1, cx, *px; if (!*out[0] || (*out[0]== ‘ ’ && *(p0[0]) == ‘ ’) || !*out[1] || (*out[1] == ‘ ’ && *(po[1]) ==‘ ’)) return; px = star; for (i = lmax+P_SPC; i; i−−) *px++ = ‘ ’; for(p0 = out[0], p1 = out[1]; *p0 && *p1; p0++, p1++) { if (isalpha(*p0) &&isalpha(*p1)) { if (xbm[*p0−‘A’]&xbm[*p1−‘A’]) { cx = ‘*’; nm++; } elseif (!dna && _day[*p0− ‘A’][*p1−‘A’] > 0) cx = ‘.’; else cx = ‘ ’; } elsecx = ‘ ’; *px++ = cx; } *px++ = ‘\n’; *px = ‘\0’; } /*  * strip path orprefix from pn, return len: pr_align()  */ static stripname(pn)stripname char *pn; /* file name (may be path) */ { register char *px,*py; py = 0; for (px = pn; *px; px++) if (*px == ‘/’) py = px + 1; if(py) (void) strcpy(pn, py); return(strlen(pn)); } /*  * cleanup() --cleanup any tmp file  * getseq() -- read in seq, set dna, len, maxlen  *g_calloc() -- calloc() with error checkin  * readjmps() -- get the goodjmps, from tmp file if necessary  * writejmps() -- write a filled arrayof jmps to a tmp file: nw()  */ #include “nw.h” #include <sys/file.h>char *jname = “/tmp/homgXXXXXX”; /* tmp file for jmps */ FILE *fj; intcleanup(); /* cleanup tmp file */ long lseek(); /*  * remove any tmpfile if we blow  */ cleanup(i) cleanup int i; { if (fj) (void)unlink(jname); exit(i); } /*  * read, return ptr to seq, set dna, len,maxlen  * skip lines starting with ‘;’, ‘<’, or ‘>’  * seq in upper orlower case  */ char * getseq(file, len) getseq char *file; /* file name*/ int *len; /* seq len */ { char line[1024], *pseq; register char *px,*py; int natgc, tlen; FILE *fp; if ((fp = fopen(file, “r”)) == 0) {fprintf(stderr, “%s: can't read %s\n”, prog, file); exit(1); } tlen =natgc = 0; while (fgets(line, 1024, fp)) { if (*line == ‘;’ || *line ==‘<’ || *line == ‘>’) continue; for (px = line; *px != ‘\n’; px++) if(isupper(*px) || islower(*px)) tlen++; } if ((pseq =malloc((unsigned)(tlen+6))) == 0) { fprintf(stderr, “%s: malloc() failedto get %d bytes for %s\n”, prog, tlen+6, file); exit(1); } pseq[0] =pseq[1] = pseq[2] = pseq[3] = ‘\0’; ...getseq py = pseq + 4; *len =tlen; rewind(fp); while (fgets(line, 1024, fp)) { if (*line == ‘;’ ||*line == ‘<’ || *line == ‘>’) continue; for (px = line; *px != ‘\n’;px++) { if (isupper(*px)) *py++ = *px; else if (islower(*px)) *py++ =toupper(*px); if (index(“ATGCU”, *(py−1))) natgc++; } } *py++ = ‘\0’;*py = ‘\0’; (void) fclose(fp); dna = natgc > (tlen/3); return(pseq+4); }char * g_calloc(msg, nx, sz) g_calloc char *msg; /* program, callingroutine */ int nx, sz; /* number and size of elements */ { char *px,*calloc(); if ((px = calloc((unsigned)nx, (unsigned)sz)) == 0) { if(*msg) { fprintf(stderr, “%s: g_calloc() failed %s (n= %d, sz= %d)\n”,prog, msg, nx, sz); exit(1); } } return(px); } /*  * get final jmps fromdx[] or tmp file, set pp[], reset dmax: main()  */ readjmps() readjmps {int fd = −1; int siz, i0, i1; register i, j, xx; if (fj) { (void)fclose(fj); if ((fd = open(jname, O_RDONLY, 0)) < 0) { fprintf(stderr,“%s: can't open() %s\n”, prog, jname); cleanup(1); } } for (i = i0 = i1= 0, dmax0 = dmax, xx = len0; ;i++) { while (1) { for (j =dx[dmax].ijmp; j >= 0 && dx[dmax].jp.x[j] >= xx; j−−) ; ...readjmps if(j < 0 && dx[dmax].offset && fj) { (void) lseek(fd, dx[dmax].offset, 0);(void) read(fd, (char *)&dx[dmax].jp, sizeof(struct jmp)); (void)read(fd, (char *)&dx[dmax].offset, sizeof(dx[dmax].offset));dx[dmax].ijmp = MAXJMP−1; } else break; } if (i >= JMPS) {fprintf(stderr, “%s: too many gaps in alignment\n”, prog); cleanup(1); }if (j >= 0) { siz = dx[dmax].jp.n[j]; xx = dx[dmax].jp.x[j]; dmax +=siz; if (siz < 0) { /* gap in second seq */ pp[1].n[il] = −siz; xx +=siz; /* id = xx − yy + len1 − 1  */ pp[1].x[il] = xx − dmax + len1 − 1;gapy++; ngapy −= siz; /* ignore MAXGAP when doing endgaps */ siz = (−siz< MAXGAP || endgaps)? −siz : MAXGAP; il++; } else if (siz > 0) { /* gapin first seq */ pp[0] .n[i0] = siz; pp[0] .x[i0] = xx; gapx++; ngapx +=siz; /* ignore MAXGAP when doing endgaps */ siz = (siz < MAXGAP ||endgaps)? siz : MAXGAP; i0++; } } else break; } /* reverse the order ofjmps  */ for (j = 0, i0−−; j < i0; j++, i0−−) { i = pp[0].n[j];pp[0].n[j] = pp[0].n[i0]; pp[0].n[i0] = i; i = pp[0].x[j]; pp[0].x[j] =pp[0].x[i0]; pp[0].x[i0] = i; } for (j = 0, i1−−; j < i1; j++, i1−−) { i= pp[1].n[j]; pp[1].n[j] = pp[1].n[i1]; pp[1].n[i1] = i; i = pp[1].x[j];pp[1].x[j] = pp[1].x[i1]; pp[1].x[i1] = i; } if (fd >= 0) (void)close(fd); if (fj) { (void) unlink(jname); fj = 0; offset = 0; } } /*  *write a filled jmp struct offset of the prev one (if any): nw()  */writejmps(ix) writejmps int ix; { char *mktemp(); if (!fj) { if(mktemp(jname) < 0) { fprintf(stderr, “%s: can't mktemp() %s\n”, prog,jname); cleanup(1); } if ((fj = fopen(jname, “w”)) == 0) {fprintf(stderr, “%s: can't write %s\n”, prog, jname); exit(1); } }(void) fwrite((char *)&dx[ix].jp, sizeof(struct jmp), 1, fj); (void)fwrite((char *)&dx[ix].offset, sizeof(dx[ix].offset), 1, fj); }

[0284] TABLE 2 PRO XXXXXXXXXXXXXXX (Length = 15 amino acids) ComparisonXXXXXYYYYYYY (Length = 12 amino acids) Protein % amino acid sequenceidentity = (the number of identically matching amino acid residuesbetween the two polypeptide sequences as determined by ALIGN-2) dividedby (the total number of amino acid residues of the PRO polypeptide) = 5divided by 15 = 33.3%

[0285] TABLE 3 PRO XXXXXXXXXX (Length = 10 amino acids) ComparisonXXXXXYYYYYYZZYZ (Length = 15 amino acids) Protein % amino acid sequenceidentity = (the number of identically matching amino acid residuesbetween the two polypeptide sequences as determined by ALIGN-2) dividedby (the total number of amino acid residues of the PRO polypeptide) = 5divided by 10 = 50%

[0286] TABLE 4 PRO-DNA NNNNNNNNNNNNNN (Length = 14 nucleotides)Comparison NNNNNNLLLLLLLLLL (Length = 16 nucleotides) DNA % nucleic acidsequence identity = (the number of identically matching nucleotidesbetween the two nucleic acid sequences as determined by ALIGN-2) dividedby (the total number of nucleotides of the PRO-DNA nucleic acidsequence) = 6 divided by 14 = 42.9%

[0287] TABLE 5 PRO-DNA NNNNNNNNNNNN (Length = 12 nucleotides) ComparisonNNNNLLLVV (Length = 9 nucleotides) DNA % nucleic acid sequence identity= (the number of identically matching nucleotides between the twonucleic acid sequences as determined by ALIGN-2) divided by (the totalnumber of nucleotides of the PRO-DNA nucleic acid sequence) = 4 dividedby 12 = 33.3%

[0288] II. Compositions and Methods of the Invention

[0289] A. Full-Length PRO Polypeptides

[0290] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO polypeptides. In particular, cDNAs encoding variousPRO polypeptides have been identified and isolated, as disclosed infurther detail in the Examples below. It is noted that proteins producedin separate expression rounds may be given different PRO numbers but theUNQ number is unique for any given DNA and the encoded protein, and willnot be changed. However, for sake of simplicity, in the presentspecification the protein encoded by the full length native nucleic acidmolecules disclosed herein as well as all further native homologues andvariants included in the foregoing definition of PRO, will be referredto as “PRO/number”, regardless of their origin or mode of preparation.

[0291] As disclosed in the Examples below, various cDNA clones have beendeposited with the ATCC. The actual nucleotide sequences of those clonescan readily be determined by the skilled artisan by sequencing of thedeposited clone using routine methods in the art. The predicted aminoacid sequence can be determined from the nucleotide sequence usingroutine skill. For the PRO polypeptides and encoding nucleic acidsdescribed herein, Applicants have identified what is believed to be thereading frame best identifiable with the sequence information availableat the time.

[0292] 1. Full-length PRO196 Polypeptides

[0293] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO196. In particular, Applicants have identified andisolated cDNAs encoding a PRO196 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that a cDNA sequence encodingfull-length native sequence PRO196 encodes for a polypeptide having anamino acid sequence which has identity with the amino acid sequence ofvarious TIE ligand polypeptides.

[0294] 2. Full-length PRO444 Polypeptides

[0295] The DNA26846-1397 clone was isolated from a human fetal lunglibrary using a trapping technique which selects for nucleotidesequences encoding secreted proteins. Thus, the DNA26846-1397 cloneencodes a secreted factor. As far as is known, the DNA26846-1397sequence encodes a novel factor designated herein as PRO444. Although,using WU-BLAST2 sequence alignment computer programs, some sequenceidentities with known proteins were revealed.

[0296] 3. Full-length PRO183 Polypeptides

[0297] The DNA28498 clone was isolated from a human tissue library. Asfar as is known, the DNA28498 sequence encodes a novel factor designatedherein as PRO183. Although, using WU-BLAST2 sequence alignment computerprograms, some sequence identities with known proteins were revealed.

[0298] 4. Full-length PRO185 Polypeptides

[0299] The DNA28503 clone was isolated from a human tissue library. Asfar as is known, the DNA28503 sequence encodes a novel factor designatedherein as PRO185. Although, using WU-BLAST2 sequence alignment computerprograms, some sequence identities with known proteins were revealed.

[0300] 5. Full-length PRO210 And PRO217 Polypeptides

[0301] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO210 and PRO217. In particular, Applicants haveidentified and isolated cDNAs encoding a PRO210 and PRO217 polypeptide,as disclosed in further detail in the Examples below. Using BLAST (FastAformat) sequence alignment computer programs, Applicants found thatcDNAs sequence encoding full-length native sequence PRO210 and PRO217have homologies to known proteins having EGF-like domains. Accordingly,it is presently believed that the PRO210 and PRO217 polypeptidesdisclosed in the present application is a newly identified member of theEGF-like family and possesses properties typical of the EGF-like proteinfamily.

[0302] 6. Full-length PRO215 Polypeptides

[0303] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO215. In particular, Applicants have identified andisolated cDNAs encoding a PRO215 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that a cDNA sequence encodingfull-length native sequence PRO215 (shown in FIG. 11 and SEQ ID NO: 16)encodes for a polypeptide having an amino acid sequence which hasidentity with the amino acid sequence of the SLIT protein precursor.PRO215 also has identity with a leucine rich repeat protein.

[0304] 7. Full-length PRO242 Polypeptides

[0305] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO242. In particular, Applicants have identified andisolated cDNA encoding a PRO242 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that a cDNA sequence encodingfull-length native sequence PRO242 (shown in FIG. 15 and SEQ ID NO: 23)has amino acid sequence identity with human macrophage inflammatoryprotein 1-alpha, rabbit macrophage inflammatory protein 1-beta, humanLD78 and rabbit immune activation gene 2. Accordingly, it is presentlybelieved that PRO242 polypeptide disclosed in the present application isa newly identified member of the chemokine family and possesses activitytypical of the chemokine family.

[0306] 8. Full-length PRO288 Polypeptides

[0307] The present invention provides newly identified and isolatedPRO288 polypeptides. In particular, Applicants have identified andisolated various human PRO288 polypeptides. The properties andcharacteristics of some of these PRO288 polypeptides are described infurther detail in the Examples below. Based upon the properties andcharacteristics of the PRO288 polypeptides disclosed herein, it isApplicants' present belief that PRO288 is a member of the TNFR family,and particularly, is a receptor for Apo-2 ligand.

[0308] 9. Full-length PRO365 Polypeptides

[0309] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO365. In particular, Applicants have identified andisolated cDNA encoding a PRO365 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO365polypeptide have significant homology with the human 2-19 protein.Accordingly, it is presently believed that PRO365 polypeptide disclosedin the present application is a newly identified member of the human2-19 protein family.

[0310] 10. Full-length PRO1361 Polypeptides

[0311] The DNA60783-1611 clone was isolated from a human B cell library.As far as is known, the DNA60783-1611 sequence encodes a novel factordesignated herein as PRO1361; using the WU-BLAST2 sequence alignmentcomputer program, no sequence identities to any known proteins wererevealed.

[0312] 11. Full-length PRO1308 Polypeptides

[0313] Using WU-BLAST2 sequence alignment computer programs, it has beenfound that PRO1308 shares certain amino acid sequence identity with theamino acid sequence of the follistatin protein designated “S55369” inthe Dayhoff database. Accordingly, it is presently believed that PRO1308disclosed in the present application is a newly identified member of thefollistatin protein family and may possess activity or propertiestypical of that family of proteins.

[0314] 12. Full-length PRO1183 Polypeptides

[0315] Using WU-BLAST2 sequence alignment computer programs, it has beenfound that a full-length native sequence PRO1183 (shown in FIG. 26 andSEQ ID NO: 52) has certain amino acid sequence identity withprotoporphyrinogen oxidase. Accordingly, it is presently believed thatPRO1183 disclosed in the present application is a newly identifiedmember of the oxidase family and may possess enzymatic activity typicalof oxidases.

[0316] 13. Full-length PRO1272 Polypeptides

[0317] Using WU-BLAST2 sequence alignment computer programs, it has beenfound that a full-length native sequence PRO1272 (shown in FIG. 28 andSEQ ID NO: 54) has certain amino acid sequence identity with cementgland-specific protein from Xenopus laevis. Accordingly, it is presentlybelieved that PRO1272 disclosed in the present application is a newlyidentified member of the XAG family and may share at least one mechanismwith the XAG proteins.

[0318] 14. Full-length PRO1419 Polypeptides

[0319] As far as is known, the DNA71290-1630 sequence encodes a novelfactor designated herein as PRO1419. Using WU-BLAST2 sequence alignmentcomputer programs, minimal sequence identities to known proteins wererevealed.

[0320] 15. Full-length PRO4999 Polypeptides

[0321] Using the ALIGN-2 sequence alignment computer program referencedabove, it has been found that the full-length native sequence PRO4999(shown in FIG. 32 and SEQ ID NO: 58) has certain amino acid sequenceidentity with UROM_HUMAN. Accordingly, it is presently believed that thePRO4999 polypeptide disclosed in the present application is a newlyidentified member of the uromodulin protein family and may possess oneor more biological and/or immunological activities or properties typicalof that protein family.

[0322] 16. Full-length PRO7170 Polypeptides

[0323] The DNA 108722-2743 clone was isolated from a human library asdescribed in the Examples below. As far as is known, the DNA 108722-2743nucleotide sequence encodes a novel factor designated herein as PRO7170;using the ALIGN-2 sequence alignment computer program, no significantsequence identities to any known proteins were revealed.

[0324] 17. Full-length PRO248 Polypeptides

[0325] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO248. In particular, Applicants have identified andisolated cDNA encoding a PRO248 polypeptide, as disclosed in furtherdetail in the Examples below. Using known programs such as BLAST andFastA sequence alignment computer programs, Applicants found that a cDNAsequence encoding full-length native sequence PRO248 (amino acidsequence shown in FIG. 36 and SEQ ID NO: 65) has certain amino acidsequence identity with growth differentiation factor 3, from mouse andfrom homo sapiens. Accordingly, it is presently believed that PRO248polypeptide disclosed in the present application is a newly identifiedmember of the transforming growth factor β family and possesses growthand differentiation capabilities typical of the this family.

[0326] 18. Full-length PRO353 Polypeptides

[0327] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO353. In particular, Applicants have identified andisolated cDNA encoding PRO353 polypeptides, as disclosed in furtherdetail in the Examples below. Using BLAST and, FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO353polypeptides have certain homology with the human and mouse complementproteins. Accordingly, it is presently believed that the PRO353polypeptides disclosed in the present application are newly identifiedmembers of the complement protein family and possesses the ability toeffect the inflammation process as is typical of the complement familyof proteins.

[0328] 19. Full-length PRO1318 and PRO1600 Polypeptides

[0329] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO1318 and PRO1600. In particular, Applicants haveidentified and isolated cDNAs encoding PRO1318 and PRO1600 polypeptides,as disclosed in further detail in the Examples below. Using BLAST andFastA sequence alignment computer programs, Applicants found that cDNAsequence encoding full-length native sequence PRO1318 and PRO1600 (shownin FIG. 40 and SEQ ID NO: 78 and FIG. 42 and SEQ ID NO: 80,respectively) have amino acid sequence identity with one or morechemokines. Accordingly, it is presently believed that the PRO1318 andPRO1600 polypeptides disclosed in the present application are newlyidentified members of the chemokine family and possesses activitytypical of the chemokine family.

[0330] 20. Full-length PRO533 Polypeptides

[0331] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO533. In particular, Applicants have identified andisolated cDNA encoding a PRO533 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST-2 and FastA sequence alignmentcomputer programs, Applicants found that a full-length native sequencePRO533 (shown in FIG. 46 and SEQ ID NO: 86) has a Blast score of 509 and53% amino acid sequence identity with fibroblast growth factor (FGF).Accordingly, it is presently believed that PRO533 disclosed in thepresent application is a newly identified member of the fibroblastgrowth factor family and may possess activity typical of suchpolypeptides.

[0332] 21. Full-length PRO301 Polypeptides

[0333] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO301. In particular, Applicants have identified andisolated cDNA encoding a PRO301 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that a full-length native sequencePRO301 (shown in FIG. 48 and SEQ ID NO: 91) has a Blast score of 246corresponding to 30% amino acid sequence identity with human A33 antigenprecursor. Accordingly, it is presently believed that PRO301 disclosedin the present application is a newly identified member of the A33antigen protein family and may be expressed in human neoplastic diseasessuch as colorectal cancer.

[0334] 22. Full-length PRO187 Polypeptides

[0335] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO187. In particular, Applicants have identified andisolated cDNA encoding a PRO187 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that a full-length native sequencePRO187 (shown in FIG. 50) has 74% amino acid sequence identity and BLASTscore of 310 with various androgen-induced growth factors and FGF-8.Accordingly, it is presently believed that PRO187 polypeptide disclosedin the present application is a newly identified member of the FGF-8protein family and may possess identify activity or property typical ofthe FGF-8-like protein family.

[0336] 23. Full-length PRO337 Polypeptides

[0337] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO337. In particular, Applicants have identified andisolated cDNA encoding a PRO337 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST, BLAST-2 and FastA sequencealignment computer programs, Applicants found that a full-length nativesequence PRO337 has 97% amino acid sequence identity with ratneurotrimin, 85% sequence identity with chicken CEPU, 73% sequenceidentity with chicken G55, 59% homology with human LAMP and 84% homologywith human OPCAM. Accordingly, it is presently believed that PRO337disclosed in the present application is a newly identified member of theIgLON sub family of the immunoglobulin superfamily and may possessneurite growth and differentiation potentiating properties.

[0338] 24. Full-length PRO1411 Polypeptides

[0339] As far as is known, the DNA59212-1627 sequence encodes a novelfactor designated herein as PRO1411. However, using WU-BLAST2 sequencealignment computer programs, some sequence identities to known proteinswere revealed.

[0340] 25. Full-length PRO4356 Polypeptides

[0341] Using WU-BLAST2 sequence alignment computer programs, it has beenfound that a full-length native sequence PRO4356 (shown in FIG. 56 andSEQ ID NO: 108) has certain amino acid sequence identity with metastasisassociated GPI-anchored protein. Accordingly, it is presently believedthat PRO4356 disclosed in the present application is a newly identifiedmember of this family and shares similar mechanisms.

[0342] 26. Full-length PRO246 Polypeptides

[0343] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO246. In particular, Applicants have identified andisolated cDNA encoding a PRO246 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that a portion of the PRO246polypeptide has significant homology with the human cell surface proteinHCAR. Accordingly, it is presently believed that PRO246 polypeptidedisclosed in the present application may be a newly identifiedmembrane-bound virus receptor or tumor cell-specific antigen.

[0344] 27. Full-length PRO265 Polypeptides

[0345] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO265. In particular, Applicants have identified andisolated cDNA encoding a PRO265 polypeptide, as disclosed in furtherdetail in the Examples below. Using programs such as BLAST and FastAsequence alignment computer programs, Applicants found that variousportions of the PRO265 polypeptide have significant homology with thefibromodulin protein and fibromodulin precursor protein. Applicants havealso found that the DNA encoding the PRO265 polypeptide has significanthomology with platelet glycoprotein V, a member of the leucine richrelated protein family involved in skin and wound repair. Accordingly,it is presently believed that PRO265 polypeptide disclosed in thepresent application is a newly identified member of the leucine richrepeat family and possesses protein protein binding capabilities, aswell as be involved in skin and wound repair as typical of this family.

[0346] 28. Full-length PRO941 Polypeptides

[0347] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO941. In particular, Applicants have identified andisolated cDNA encoding a PRO941 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO941 polypeptide hassignificant similarity to one or more cadherin proteins. Accordingly, itis presently believed that PRO941 polypeptide disclosed in the presentapplication is a newly identified cadherin homolog.

[0348] 29. Full-length PRO10096 polypeptides

[0349] Using the ALIGN-2 sequence alignment computer program referencedabove, it has been found that the full-length native sequence PRO10096(shown in FIG. 64 and SEQ ID NO: 126) has certain amino acid sequenceidentity with various interleukin-10-related molecules. Accordingly, itis presently believed that the PRO10096 polypeptide disclosed in thepresent application is a newly identified IL-10 homolog and may possessone or more biological and/or immunological activities or propertiestypical of that protein.

[0350] 30. Full-length PRO6003 Polypeptides

[0351] The DNA83568-2692 clone was isolated from a human fetal kidneylibrary as described in the Examples below. As far as is known, theDNA83568-2692 nucleotide sequence encodes a novel factor designatedherein as PRO6003; using the ALIGN-2 sequence alignment computerprogram, no significant sequence identities to any known proteins wererevealed.

[0352] B. PRO Polypeptide Variants

[0353] In addition to the full-length native sequence PRO polypeptidesdescribed herein, it is contemplated that PRO variants can be prepared.PRO variants can be prepared by introducing appropriate nucleotidechanges into the PRO DNA, and/or by synthesis of the desired PROpolypeptide. Those skilled in the art will appreciate that amino acidchanges may alter post-translational processes of the PRO, such aschanging the number or position of glycosylation sites or altering themembrane anchoring characteristics.

[0354] Variations in the native full-length sequence PRO or in variousdomains of the PRO described herein, can be made, for example, using anyof the techniques and guidelines for conservative and non-conservativemutations set forth, for instance, in U.S. Pat. No. 5,364,934.Variations may be a substitution, deletion or insertion of one or morecodons encoding the PRO that results in a change in the amino acidsequence of the PRO as compared with the native sequence PRO. Optionallythe variation is by substitution of at least one amino acid with anyother amino acid in one or more of the domains of the PRO. Guidance indetermining which amino acid residue may be inserted, substituted ordeleted without adversely affecting the desired activity may be found bycomparing the sequence of the PRO with that of homologous known proteinmolecules and minimizing the number of amino acid sequence changes madein regions of high homology. Amino acid substitutions can be the resultof replacing one amino acid with another amino acid having similarstructural and/or chemical properties, such as the replacement of aleucine with a scrine, i.e., conservative amino acid replacements.Insertions or deletions may optionally be in the range of about 1 to 5amino acids. The variation allowed may be determined by systematicallymaking insertions, deletions or substitutions of amino acids in thesequence and testing the resulting variants for activity exhibited bythe full-length or mature native sequence.

[0355] PRO polypeptide fragments are provided herein. Such fragments maybe truncated at the N-terminus or C-terminus, or may lack internalresidues, for example, when compared with a full length native protein.Certain fragments lack amino acid residues that are not essential for adesired biological activity of the PRO polypeptide.

[0356] PRO fragments may be prepared by any of a number of conventionaltechniques. Desired peptide fragments may be chemically synthesized. Analternative approach involves generating PRO fragments by enzymaticdigestion, e.g., by treating the protein with an enzyme known to cleaveproteins at sites defined by particular amino acid residues, or bydigesting the DNA with suitable restriction enzymes and isolating thedesired fragment. Yet another suitable technique involves isolating andamplifying a DNA fragment encoding a desired polypeptide fragment, bypolymerase chain reaction (PCR). Oligonucleotides that define thedesired termini of the DNA fragment are employed at the 5′ and 3′primers in the PCR. Preferably, PRO polypeptide fragments share at leastone biological and/or immunological activity with the native PROpolypeptide disclosed herein.

[0357] In particular embodiments, conservative substitutions of interestare shown in Table 6 under the heading of preferred substitutions. Ifsuch substitutions result in a change in biological activity, then moresubstantial changes, denominated exemplary substitutions in Table 6, oras further described below in reference to amino acid classes, areintroduced and the products screened. TABLE 6 Original ExemplaryPreferred Residue Substitutions Substitutions Ala (A) val; leu; ile valArg (R) lys; gln; asn lys Asn (N) gln; his; lys; arg gln Asp (D) glu gluCys (C) ser ser Gln (Q) asn asn Glu (E) asp asp Gly (G) pro; ala ala His(H) asn; gln; lys; arg arg Ile (I) leu; val; met; ala; phe; norleucineleu Leu (L) norleucine; ile; val; met; ala; phe ile Lys (K) arg; gln;asn arg Met (M) leu; phe; ile leu Phe (F) leu; val; ile; ala; tyr leuPro (P) ala ala Ser (S) thr thr Thr (T) ser ser Trp (W) tyr; phe tyr Tyr(Y) trp; phe; thr; ser phe Val (V) ile; leu; met; phe; ala; norleucineleu

[0358] Substantial modifications in function or immunological identityof the PRO polypeptide are accomplished by selecting substitutions thatdiffer significantly in their effect on maintaining (a) the structure ofthe polypeptide backbone in the area of the substitution, for example,as a sheet or helical conformation, (b) the charge or hydrophobicity ofthe molecule at the target site, or (c) the bulk of the side chain.Naturally occurring residues are divided into groups based on commonside-chain properties:

[0359] (1) hydrophobic: norleucine, met, ala, val, leu, ile;

[0360] (2) neutral hydrophilic: cys, ser, thr;

[0361] (3) acidic: asp, glu;

[0362] (4) basic: asn, gln, his, lys, arg;

[0363] (5) residues that influence chain orientation: gly, pro; and

[0364] (6) aromatic: trp, tyr, phe.

[0365] Non-conservative substitutions will entail exchanging a member ofone of these classes for another class. Such substituted residues alsomay be introduced into the conservative substitution sites or, morepreferably, into the remaining (non-conserved) sites.

[0366] The variations can be made using methods known in the art such asoligonucleotide-mediated (site-directed) mutagenesis, alanine scanning,and PCR mutagenesis. Site-directed mutagenesis [Carter et al., Nucl.Acids Res., 13:4331 (1986); Zoller et al., Nucl. Acids Res., 10:6487(1987)], cassette mutagenesis [Wells et al., Gene, 34:315 (1985)],restriction selection mutagenesis [Wells et al., Philos. Trans. R. Soc.London SerA, 317:415 (1986)] or other known techniques can be performedon the cloned DNA to produce the PRO variant DNA.

[0367] Scanning amino acid analysis can also be employed to identify oneor more amino acids along a contiguous sequence. Among the preferredscanning amino acids are relatively small, neutral amino acids. Suchamino acids include alanine, glycine, serine, and cysteine. Alanine istypically a preferred scanning amino acid among this group because iteliminates the side-chain beyond the beta-carbon and is less likely toalter the main-chain conformation of the variant [Cunningham and Wells,Science, 244: 1081-1085 (1989)]. Alanine is also typically preferredbecause it is the most common amino acid. Further, it is frequentlyfound in both buried and exposed positions [Creighton, The Proteins, (W.H. Freeman & Co., N.Y.); Chothia, J. Mol. Biol., 150:1 (1976)]. Ifalanine substitution does not yield adequate amounts of variant, anisoteric amino acid can be used.

[0368] C. Modifications of PRO

[0369] Covalent modifications of PRO are included within the scope ofthis invention. One type of covalent modification includes reactingtargeted amino acid residues of a PRO polypeptide with an organicderivatizing agent that is capable of reacting with selected side chainsor the N- or C- terminal residues of the PRO. Derivatization withbifunctional agents is useful, for instance, for crosslinking PRO to awater-insoluble support matrix or surface for use in the method forpurifying anti-PRO antibodies, and vice-versa. Commonly usedcrosslinking agents include, e.g., 1,1-bis(diazoacetyl)-2-phenylethlane,glutaraldehyde, N-hydroxysuccinimide esters, for example, esters with4-azidosalicylic acid, homobifunctional imidoesters, includingdisuccinimidyl esters such as 3,3′-dithiobis(succinimidylpropionate),bifunctional maleimides such as bis-N-maleimido-1,8-octane and agentssuch as methyl-3-[(p-azidophenyl)dithio]propioimidate.

[0370] Other modifications include deamidation of glutaminyl andasparaginyl residues to the corresponding glutamyl and aspartylresidues, respectively, hydroxylation of proline and lysine,phosphorylation of hydroxyl groups of seryl or threonyl residues,methylation of the α-amino groups of lysine, arginine, and histidineside chains [T. E. Creighton, Proteins: Structure and MolecularProperties, W. H. Freeman & Co., San Francisco, pp. 79-86 (1983)],acetylation of the N-terminal amine, and amidation of any C-terminalcarboxyl group.

[0371] Another type of covalent modification of the PRO polypeptideincluded within die scope of this invention comprises altering thenative glycosylation pattern of the polypeptide. “Altering the nativeglycosylation pattern” is intended for purposes herein to mean deletingone or more carbohydrate moieties found in native sequence PRO (eitherby removing the underlying glycosylation site or by deleting theglycosylation by chemical and/or enzymatic means), and/or adding one ormore glycosylation sites that are not present in the native sequencePRO. In addition, the phrase includes qualitative changes in theglycosylation of the native proteins, involving a change in the natureand proportions of the various carbohydrate moieties present.

[0372] Addition of glycosylation sites to the PRO polypeptide may beaccomplished by altering the amino acid sequence. The alteration may bemade, for example, by the addition of, or substitution by, one or moreserine or threonine residues to the native sequence PRO (for O-linkedglycosylation sites). The PRO amino acid sequence may optionally bealtered through changes at the DNA level, particularly by mutating theDNA encoding the PRO polypeptide at preselected bases such that codonsare generated that will translate into the desired amino acids.

[0373] Another means of increasing the number of carbohydrate moietieson the PRO polypeptide is by chemical or enzymatic coupling ofglycosides to the polypeptide. Such methods are described in the art,e.g., in WO 87/05330 published Sep. 11, 1987, and in Aplin and Wriston,CRC Crit. Rev. Biochem., pp. 259-306 (1981).

[0374] Removal of carbohydrate moieties present on the PRO polypeptidemay be accomplished chemically or enzymatically or by mutationalsubstitution of codons encoding for amino acid residues that serve astargets for glycosylation. Chemical deglycosylation techniques are knownin the art and described, for instance, by Hakimuddin, et al., Arch.Biochem. Biophvs., 259:52 (1987) and by Edge et al., Anal. Biochem.,118:131 (1981). Enzymatic cleavage of carbohydrate moieties onpolypeptides can be achieved by the use of a variety of endo- andexo-glycosidases as described by Thotakura et al., Meth. Enzymol.,138:350 (1987).

[0375] Another type of covalent modification of PRO comprises linkingthe PRO polypeptide to one of a variety of nonproteinaceous polymers,e.g., polyethylene glycol (PEG), polypropylene glycol, orpolyoxyalkylenes, in the manner set forth in U.S. Pat. Nos. 4,640,835;4,496,689; 4,301,144; 4,670,417; 4,791,192 or 4,179,337.

[0376] The PRO of the present invention may also be modified in a way toform a chimeric molecule comprising PRO fused to another, heterologouspolypeptide or amino acid sequence.

[0377] In one embodiment, such a chimeric molecule comprises a fusion ofthe PRO with a tag polypeptide which provides an epitope to which ananti-tag antibody can selectively bind. The epitope tag is generallyplaced at the amino- or carboxyl-terminus of the PRO. The presence ofsuch epitope-tagged forms of the PRO can be detected using an antibodyagainst the tag polypeptide. Also, provision of the epitope tag enablesthe PRO to be readily purified by affinity purification using ananti-tag antibody or another type of affinity matrix that binds to theepitope tag. Various tag polypeptides and their respective antibodiesare well known in the art. Examples include poly-histidine (poly-his) orpoly-histidine-glycine (poly-his-gly) tags; the flu HA tag polypeptideand its antibody 12CA5 [Field et al., Mol. Cell. Biol., 8:2159-2165(1988)]; the c-myc tag and the 8F9, 3C7, 6E10, G4, B7 and 9E10antibodies thereto [Evan et al., Molecular and Cellular Biology,5:3610-3616 (1985)]; and the Herpes Simplex virus glycoprotein D (gD)tag and its antibody [Paborsky et al., Protein Engineering, 3(6):547-553(1990)]. Other tag polypeptides include the Flag-peptide [Hopp et al.,BioTechnology, 6:1204-1210 (1988)]; the KT3 epitope peptide [Martin etal., Science, 255:192-194 (1992); an α-tubulin epitope peptide [Skinneret al., J. Biol. Chem, 266:15163-15166 (1991)]; and the T7 gene 10protein peptide tag [Lutz-Freyermuth et al., Proc. Natl. Acad. Sci. USA,87:6393-6397 (1990)].

[0378] In an alternative embodiment, the chimeric molecule may comprisea fusion of the PRO with an immunoglobulin or a particular region of animmunoglobulin. For a bivalent form of the chimeric molecule (alsoreferred to as an “immunoadhesin”), such a fusion could be to the Feregion of an IgG molecule. The Ig fusions preferably include thesubstitution of a soluble (transmembrane domain deleted or inactivated)form of a PRO polypeptide in place of at least one variable regionwithin an Ig molecule. In a particularly preferred embodiment, theimmunoglobulin fusion includes the hinge, CH2 and CH3, or the hinge,CH1, CH2 and CH3 regions of an IgGl molecule. For the production ofimmunoglobulin fusions see also U.S Pat. No. 5,428,130 issued Jun. 27,1995.

[0379] D. Preparation of PRO

[0380] The description below relates primarily to production of PRO byculturing cells transformed or transfected with a vector containing PROnucleic acid. It is, of course, contemplated that alternative methods,which are well known in the art, may be employed to prepare PRO. Forinstance, the PRO sequence, or portions thereof, may be produced bydirect peptide synthesis using solid-phase techniques [see, e.g.,Stewart et al., Solid-Phase Peptide Synthesis, W. H. Freeman Co., SanFrancisco, Calif. (1969); Merrifield, J. Am. Chem. Soc., 85:2149-2154(1963)]. In vitro protein synthesis may be performed using manualtechniques or by automation. Automated synthesis may be accomplished,for instance, using an Applied Biosystems Peptide Synthesizer (FosterCity, Clif.) using manufacturer's instructions. Various portions of thePRO may be chemically synthesized separately and combined using chemicalor enzymatic methods to produce the full-length PRO.

[0381] 1. Isolation of DNA Encoding PRO

[0382] DNA encoding PRO may be obtained from a cDNA library preparedfrom tissue believed to possess the PRO mRNA and to express it at adetectable level. Accordingly, human PRO DNA can be convenientlyobtained from a cDNA library prepared from human tissue, such asdescribed in the Examples. The PRO-encoding gene may also be obtainedfrom a genomic library or by known synthetic procedures (e.g., automatednucleic acid synthesis).

[0383] Libraries can be screened with probes (such as antibodies to thePRO or oligonucleotides of at least about 20-80 bases) designed toidentify the gene of interest or the protein encoded by it. Screeningthe cDNA or genomic library with the selected probe may be conductedusing standard procedures, such as described in Sambrooketal., MolecularCloning: A Laboratory Manual (New York: Cold Spring Harbor LaboratoryPress, 1989). An alternative means to isolate the gene encoding PRO isto use PCR methodology [Sambrook et al., supra; Dieffenbach et al., PCRPrimer: A Laboratory Manual (Cold Spring Harbor Laboratory Press,1995)].

[0384] The Examples below describe techniques for screening a cDNAlibrary. The oligonucleotide sequences selected as probes should be ofsufficient length and sufficiently unambiguous that false positives areminimized. The oligonucleotide is preferably labeled such that it can bedetected upon hybridization to DNA in the library being screened.Methods of labeling are well known in the art, and include the use ofradiolabels like ³²P-labeled ATP, biotinylation or enzyme labeling.Hybridization conditions, including moderate stringency and highstringency, are provided in Sambrook et al., supra.

[0385] Sequences identified in such library screening methods can becompared and aligned to other known sequences deposited and available inpublic databases such as GenBank or other private sequence databases.Sequence identity (at either the amino acid or nucleotide level) withindefined regions of the molecule or across the full-length sequence canbe determined using methods known in the art and as described herein.

[0386] Nucleic acid having protein coding sequence may be obtained byscreening selected cDNA or genomic libraries using the deduced aminoacid sequence disclosed herein for the first time, and, if necessary,using conventional primer extension procedures as described in Sambrooket al., supra, to detect precursors and processing intermediates of mRNAthat may not have been reverse-transcribed into cDNA.

[0387] 2. Selection and Transformation of Host Cells

[0388] Host cells are transfected or transformed with expression orcloning vectors described herein for PRO production and cultured inconventional nutrient media modified as appropriate for inducingpromoters, selecting transformants, or amplifying the genes encoding thedesired sequences. The culture conditions, such as media, temperature,pH and the like, can be selected bv the skilled artisan without undueexperimentation. In general, principles, protocols, and practicaltechniques for maximizing the productivity of cell cultures can be foundin Mammalian Cell Biotechnology: a Practical Approach, M. Butler, ed.(IRL Press, 1991) and Sambrook et al., supra.

[0389] Methods of eukaryotic cell transfection and prokaryotic celltransformation are known to the ordinarily skilled artisan, for example,CaCl₂, CaPO₄, liposome-mediated and electroporation. Depending on thehost cell used, transformation is performed using standard techniquesappropriate to such cells. The calcium treatment employing calciumchloride, as described in Sambrook et al., supra, or electroporation isgenerally used for prokaryotes. Infection with Agrobacterium tumefaciensis used for transformation of certain plant cells, as described by Shawet al., Gene, 23:315 (1983) and WO 89/05859 published Jun. 29, 1989. Formammalian cells without such cell walls, the calcium phosphateprecipitation method of Graham and van der Eb, Virology, 52:456-457(1978) can be employed. General aspects of mammalian cell host systemtransfections have been described in U.S. Pat. No. 4,399,216.Transformations into yeast are typically carried out according to themethod of Van Solingen et al., J. Bact., 130:946 (1977) and Hsiao etal., Proc. Natl. Acad. Sci. (USA), 76:3829 (1979). However, othermethods for introducing DNA into cells, such as by nuclearmicroinjection, electroporation, bacterial protoplast fusion with intactcells, or polycations, e.g., polybrene, polyornithine, may also be used.For various techniques for transforming mammalian cells, see Keown etal., Methods in Enzymology, 185:527-537 (1990) and Mansour et al.,Nature, 336:348-352 (1988).

[0390] Suitable host cells for cloning or expressing the DNA in thevectors herein include prokaryote, yeast, or higher eukaryote cells.Suitable prokaryotes include but are not limited to eubacteria, such asGram-negative or Gram-positive organisms, for example,Enterobacteriaceae such as E. coli. Various E. coli strains are publiclyavailable, such as E. coli K12 strain MM294 (ATCC 31,446); E. coli X1776(ATCC 31,537); E. coli strain W3110 (ATCC 27,325) and K5 772 (ATCC53,635). Other suitable prokaryotic host cells includeEnterobacteriaceae such as Escherichia, e.g., E. coli, Enterobacter,Erwinia, Klebsiella, Proteus, Salmonella, e.g., Salmonella typhimurium,Serratia, e.g., Serratia marcescans, and Shigella, as well as Bacillisuch as B. subtilis and B. licheniformis (e.g., B. licheniformis 41Pdisclosed in DD 266,710 published Apr. 12, 1989), Pseudomonas such as P.Aeruginosa, and Streptomyces. These examples are illustrative ratherthan limiting. Strain W3110 is one particularly preferred host or parenthost because it is a common host strain for recombinant DNA productfermentations. Preferably, the host cell secretes minimal amounts ofproteolytic enzymes. For example, strain W3110 may be modified to effecta genetic mutation in the genes encoding proteins endogenous to thehost, with examples of such hosts including E. coli W3110 strain IA2,which has the complete genotype tonA ; E. coli W3110 strain 9E4, whichhas the complete genotype tonA ptr3; E. coli W3110 strain 27C7 (ATCC55,244), which has the complete genotype tonA ptr3 phoA E15(argF-lac)169 degP onpT kan'; E. coli W3110 strain 37D6, which has thecomplete genotype tonA ptr3 phoA E15 (argF-lac)169 degP ompT rbs7 ilvGkan'; E. coli W3110 strain 40B4, which is strain 37D6 with anon-kanamycin resistant degP deletion mutation; and an E. coli strainhaving mutant periplasmic protease disclosed in U.S. Pat. No. 4,946,783issued Aug. 7, 1990. Alternatively, in vitro methods of cloning, e.g.,PCR or other nucleic acid polymerase reactions, are suitable.

[0391] In addition to prokaryotes, eukaryotic microbes such asfilamentous fungi or yeast are suitable cloning or expression hosts forPRO-encoding vectors. Saccharomyces cerevisiae is a commonly used lowereukaryotic host microorganism. Others include Schizosaccharomyces pombe(Beach and Nurse, Nature, 290: 140 [1981]; EP 139,383 published May 2,1985); Kluyveromyces hosts (U.S. Pat. No. 4,943,529; Fleer et al.,Bio/Technology, 9:968-975 (1991)) such as, e.g., K. lactis (MW98-8C,CBS683, CBS4574; Louvencourt et al., J. Bacteriol., 154(2):737-742[1983]), K. fragilis (ATCC 12,424), K. bulgaricus (ATCC 16,045), K.wickeramii (ATCC 24,178), K. waltii (ATCC 56,500), K. drosophilarum(ATCC 36,906; Van den Berg et al., Bio/Technology, 8:135 (1990)), K.thermotolerans, and K. mairxianus; yarroivia (EP 402,226); Pichiapastoris (EP 183,070; Sreekrishna et al., J. Basic Microbiol.,28:265-278 [1988]); Candida; Trichoderma reesia (EP 244,234); Neurosporacrassa (Case et al., Proc. NatI. Acad. Sci. USA, 76:5259-5263 [1979]);Schwanniomnyces such as Schwanniomyces occidentalis (EP 394,538published Oct. 31, 1990); and filamentous fungi such as, e.g.,Neurospora, Penicillium, Tolypocladiuni (WO 91/00357 published Jan. 10,1991), and Aspergillus hosts such as A. nidulans (Allance et al .,Biochem. Biophys. Res. Commun., 112:284-289 [1983]; Tilburn et al.,Gene, 26:205-221 [1983]; Yelton et al., Proc. Natl. Acad. Sci. USA,81:14-1474 [1984]) and A. niger (Kelly and Hynes, EMBO J., 4:475-479[1985]). Methylotropic yeasts are suitable herein and include, but arenot limited to, yeast capable of growth on methanol selected from thegenera consisting of Hansenula, Candida, Kloeckera, Pichia,Saccharomyces, Torulopsis, and Rhodotorula. A list of specific speciesthat are exemplary of this class of yeasts may be found in C. Anthony,The Biochemistry of Methylotrophs, 269 (1982).

[0392] Suitable host cells for the expression of glycosylated PRO arederived from multicellular organisms. Examples of invertebrate cellsinclude insect cells such as Drosophila S2 and Spodoptera Sf9, as wellas plant cells. Examples of useful mammalian host cell lines includeChinese hamster ovary (CHO) and COS cells. More specific examplesinclude monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL1651); human embryonic kidney line (293 or 293 cells subcloned forgrowth in suspension culture, Graham et al., J. Gen Virol., 36:59(1977)); Chinese hamster ovary cells/-DHFR (CHO, Urlaub and Chasin,Proc. Natl. Acad. Sci. USA, 77:4216 (1980)); mouse sertoli cells (TM4,Mather, Biol. Reprod., 23:243-251 (1980)); human lung cells (W138, ATCCCCL 75); human liver cells (Hep G2, HB 8065); and mouse mammary tumor(MMT 060562, ATCC CCL51). The selection of the appropriate host cell isdeemed to be within the skill in the art.

[0393] 3. Selection and Use of a Replicable Vector

[0394] The nucleic acid (e.g., cDNA or genomic DNA) encoding PRO may beinserted into a replicable vector for cloning (amplification of the DNA)or for expression. Various vectors are publicly available. The vectormay, for example, be in the form of a plasmid, cosmid, viral particle,or phage. The appropriate nucleic acid sequence may be inserted into thevector by a variety of procedures. In general, DNA is inserted into anappropriate restriction endonuclease site(s) using techniques known inthe art. Vector components generally include, but are not limited to,one or more of a signal sequence, an origin of replication, one or moremarker genes, an enhancer element, a promoter, and a transcriptiontermination sequence. Construction of suitable vectors containing one ormore of these components employs standard ligation techniques which areknown to the skilled artisan.

[0395] The PRO may be produced recombinantly not only directly, but alsoas a fusion polypeptide with a heterologous polypeptide, which may be asignal sequence or other polypeptide having a specific cleavage site atthe N-terminus of the mature protein or polypeptide. In general, thesignal sequence may be a component of the vector, or it may be a part ofthe PRO-encoding DNA that is inserted into the vector. The signalsequence may be a prokaryotic signal sequence selected, for example,from the group of the alkaline phosphatase, penicillinase, lpp, orheat-stable enterotoxin II leaders. For yeast secretion the signalsequence may be, e.g., the yeast invertase leader, alpha factor leader(including Saccharomyces and Kluyveroinyces α-factor leaders, the latterdescribed in U.S. Pat. No. 5,010,182), or acid phosphatase leader, theC. albicans glucoamylase leader (EP 362,179 published Apr. 4, 1990), orthe signal described in WO 90/13646 published Nov. 15, 1990. Inmammalian cell expression, mammalian signal sequences may be used todirect secretion of the protein, such as signal sequences from secretedpolypeptides of the same or related species, as well as viral secretoryleaders.

[0396] Both expression and cloning vectors contain a nucleic acidsequence that enables the vector to replicate in one or more selectedhost cells. Such sequences are well known for a variety of bacteria,yeast, and viruses. The origin of replication from the plasmid pBR322 issuitable for most Gram-negative bacteria, the 2μ plasmid origin issuitable for yeast, and various viral origins (SV40, polyoma,adenovirus, VSV or BPV) are useful for cloning vectors in mammaliancells.

[0397] Expression and cloning vectors will typically contain a selectiongene, also termed a selectable marker. Typical selection genes encodeproteins that (a) confer resistance to antibiotics or other toxins,e.g., ampicillin, neomycin, methotrexate, or tetracycline, (b)complement auxotrophic deficiencies, or (c) supply critical nutrientsnot available from complex media, e.g., the gene encoding D-alanineracemase for Bacilli.

[0398] An example of suitable selectable markers for mammalian cells arethose that enable the identification of cells competent to take up thePRO-encoding nucleic acid, such as DHFR or thymidine kinase. Anappropriate host cell when wild-type DHFR is employed is the CHO cellline deficient in DHFR activity, prepared and propagated as described byUrlaub et al., Proc. Natl. Acad. Sci. USA, 77:4216 (1980). A suitableselection gene for use in yeast is the trp1 gene present in the yeastplasmid YRp7 [Stinchcomb et al., Nature, 282:39 (1979); Kingsman et al.,Gene, 7:141 (1979); Tschemper et al., Gene, 10:157 (1980)]. The trp1gene provides a selection marker for a mutant strain of yeast lackingthe ability to grow in tryptophan, for example, ATCC No. 44076 or PEP4-1[Jones, Genetics, 85:12 (1977)].

[0399] Expression and cloning vectors usually contain a promoteroperably linked to the PRO-encoding nucleic acid sequence to direct mRNAsynthesis. Promoters recognized by a variety of potential host cells arewell known. Promoters suitable for use with prokaryotic hosts includethe β-lactamase and lactose promoter systems [Chang et al., Nature,275:615 (1978); Goeddel et al., Nature, 281:544 (1979)], alkalinephosphatase, a tryptophan (trp) promoter system [Goeddel, Nucleic AcidsRes., 8:4057 (1980); EP 36,776], and hybrid promoters such as the tacpromoter [deBoer et al., Proc. Natl. Acad. Sci. USA, 80:21-25 (1983)].Promoters for use in bacterial systems also will contain aShine-Dalgarno (S. D.) sequence operably linked to the DNA encoding PRO.

[0400] Examples of suitable promoting sequences for use with yeast hostsinclude the promoters for 3-phosphoglycerate kinase [Hitzeman et al., J.Biol. Chem., 255:2073(1980)] or other glycolytic enzymes [Hess et al.,J. Adv. Enzyme Reg., 7:149 (1968); Holland, Biochemistry, 17:4900(1978)], such as enolase, glyceraldehyde-3-phosphate dehydrogenase,hexokinase, pyruvate decarboxylase, phosphofructokinase,glucose-6-phosphate isomerase, 3-phosphoglycerate mutase, pyruvatekinase, triosephosphate isomerase, phosphoglucose isomerase, andglucokinase.

[0401] Other yeast promoters, which are inducible promoters having theadditional advantage of transcription controlled by growth conditions,are the promoter regions for alcohol dehydrogenase 2, isocytochrome C,acid phosphatase, degradative enzymes associated with nitrogenmetabolism, metallothionein, glyceraldehyde-3-phosphate dehydrogenase,and enzymes responsible for maltose and galactose utilization. Suitablevectors and promoters for use in yeast expression are further describedin EP 73,657.

[0402] PRO transcription from vectors in mammalian host cells iscontrolled, for example, by promoters obtained from the genomes ofviruses such as polyoma virus, fowlpox virus (UK 2,211,504 publishedJul. 5, 1989), adenovirus (such as Adenovirus 2), bovine papillomavirus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-Bvirus and Simian Virus 40 (SV40), from heterologous mammalian promoters,e.g., the actin promoter or an immunoglobulin promoter, and fromheat-shock promoters, provided such promoters are compatible with thehost cell systems.

[0403] Transcription of a DNA encoding the PRO by higher eukaryotes maybe increased by inserting an enhancer sequence into the vector.Enhancers are cis-acting elements of DNA, usually about from 10 to 300bp, that act on a promoter to increase its transcription. Many enhancersequences are now known from mammalian genes (globin, elastase, albumin,a-fetoprotein, and insulin). Typically, however, one will use anenhancer from a eukaryotic-cell virus. Examples include the SV40enhancer on the late side of the replication origin (bp 100-270), thecytomegalovirus early promoter enhancer, the polyoma enhancer on thelate side of the replication origin, and adenovirus enhancers. Theenhancer may be spliced into the vector at a position 5′ or 3′ to thePRO coding sequence, but is preferably located at a site 5′ from thepromoter.

[0404] Expression vectors used in eukaryotic host cells (yeast, fungi,insect, plant, animal, human, or nucleated cells from othermulticellular organisms) will also contain sequences necessary for thetermination of transcription and for stabilizing the mRNA. Suchsequences are commonly available from the 5′ and, occasionally 3′,untranslated regions of eukaryotic or viral DNAs or cDNAs. These regionscontain nucleotide segments transcribed as polyadenylated fragments inthe untranslated portion of the mRNA encoding PRO.

[0405] Still other methods, vectors, and host cells suitable foradaptation to the synthesis of PRO in recombinant vertebrate cellculture are described in Gething et al., Nature, 293:620-625 (1981);Mantei et al., Nature, 281:40-46 (1979); EP 117,060; and EP 117,058.

[0406]4. Detecting Gene Amplification/Expression

[0407] Gene amplification and/or expression may be measured in a sampledirectly, for example, by conventional Southern blotting, Northernblotting to quantitate the transcription of mRNA [Thomas, Proc. Natl.Acad. Sci. USA, 77:5201-5205 (1980)], dot blotting (DNA analysis), or insitu hybridization, using an appropriately labeled probe, based on thesequences provided herein. Alternatively, antibodies may be employedthat can recognize specific duplexes, including DNA duplexes, RNAduplexes, and DNA-RNA hybrid duplexes or DNA-protein duplexes. Theantibodies in turn may be labeled and the assay may be carried out wherethe duplex is bound to a surface, so that upon the formation of duplexon the surface, the presence of antibody bound to the duplex can bedetected.

[0408] Gene expression, alternatively, may be measured by immunologicalmethods, such as immunohistochemical staining of cells or tissuesections and assay of cell culture or body fluids, to quantitatedirectly the expression of gene product. Antibodies useful forimmunohistoclhemical staining and/or assay of sample fluids may beeither monoclonal or polyclonal, and may be prepared in any mammal.Conveniently, the antibodies may be prepared against a native sequencePRO polypeptide or against a synthetic peptide based on the DNAsequences provided herein or against exogenous sequence fused to PRO DNAand encoding a specific antibody epitope.

[0409] 5. Purification of Polypeptide

[0410] Forms of PRO may be recovered from culture medium or from hostcell lysates. If membrane-bound, it can be released from the membraneusing a suitable detergent solution (e.g. Triton-X 100) or by enzymaticcleavage. Cells employed in expression of PRO can be disrupted byvarious physical or chemical means, such as freeze-thaw cycling,sonication, mechanical disruption, or cell lysing agents.

[0411] It may be desired to purify PRO from recombinant cell proteins orpolypeptides. The following procedures are exemplary of suitablepurification procedures: by fractionation on an ion-exchange column;ethanol precipitation; reverse phase HPLC; chromatography on silica oron a cation-exchange resin such as DEAE; chromatofocusing; SDS-PAGE;ammonium sulfate precipitation; gel filtration using, for example,Sephadex G-75; protein A Sepharose columns to remove contaminants suchas IgG; and metal chelating columns to bind epitope-tagged forms of thePRO. Various methods of protein purification may be employed and suchmethods are known in the art and described for example in Deutscher,Methods in Enzymology, 182 (1990); Scopes, Protein Purification:Principles and Practice, Springer-Verlag, New York (1982). Thepurification step(s) selected will depend, for example, on the nature ofthe production process used and the particular PRO produced.

[0412] E. Uses for PRO

[0413] Nucleotide sequences (or their complement) encoding PRO havevarious applications in the art of molecular biology, including uses ashybridization probes, in chromosome and gene mapping and in thegeneration of anti-sense RNA and DNA. PRO nucleic acid will also beuseful for the preparation of PRO polypeptides by the recombinanttechniques described herein.

[0414] The full-length native sequence PRO gene, or portions thereof,may be used as hybridization probes for a cDNA library to isolate thefull-length PRO cDNA or to isolate still other cDNAs (for instance,those encoding naturally-occurring variants of PRO or PRO from otherspecies) which have a desired sequence identity to the native PROsequence disclosed herein. Optionally, the length of the probes will beabout 20 to about 50 bases. The hybridization probes may be derived fromat least partially novel regions of the full length native nucleotidesequence wherein those regions may be determined without undueexperimentation or from genomic sequences including promoters, enhancerelements and introns of native sequence PRO. By way of example, ascreening method will comprise isolating the coding region of the PROgene using the known DNA sequence to synthesize a selected probe ofabout 40 bases. Hybridization probes may be labeled by a variety oflabels, including radionucleotides such as ³²P or ³S, or enzymaticlabels such as alkaline phosphatase coupled to the probe viaavidin/biotin coupling systems. Labeled probes having a sequencecomplementary to that of the PRO gene of the present invention can beused to screen libraries of human cDNA, genomic DNA or mRNA to determinewhich members of such libraries the probe hybridizes to. Hybridizationtechniques are described in further detail in the Examples below.

[0415] Any EST sequences disclosed in the present application maysimilarly be employed as probes, using the methods disclosed herein.

[0416] Other useful fragments of the PRO nucleic acids include antisenseor sense oligonucleotides comprising a singe-stranded nucleic acidsequence (either RNA or DNA) capable of binding to target PRO mRNA(sense) or PRO DNA (antisense) sequences. Antisense or senseoligonucleotides, according to the present invention, comprise afragment of the coding region of PRO DNA. Such a fragment generallycomprises at least about 14 nucleotides, preferably from about 14 to 30nucleotides. The ability to derive an antisense or a senseoligonucleotide, based upon a cDNA sequence encoding a given protein isdescribed in, for example, Stein and Cohen (Cancer Res. 48:2659, 1988)and van der Krol et al. (BioTechniques 6:958, 1988).

[0417] Binding of antisense or sense oligonucleotides to target nucleicacid sequences results in the formation of duplexes that blocktranscription or translation of the target sequence by one of severalmeans, including enhanced degradation of the duplexes, prematuretermination of transcription or translation, or by other means. Theantisense oligonucleotides thus may be used to block expression of PROproteins. Antisense or sense oligonucleotides further compriseoligonucleotides having modified sugar-phosphodiester backbones (orother sugar linkages, such as those described in WO 91/06629) andwherein such sugar linkages are resistant to endogenous nucleases. Sucholigonucleotides with resistant sugar linkages are stable in vivo (i.e.,capable of resisting enzymatic degradation) but retain sequencespecificity to be able to bind to target nucleotide sequences.

[0418] Other examples of sense or antisense oligonucleotides includethose oligonucleotides which are covalently linked to organic moieties,such as those described in WO 90/10048, and other moieties thatincreases affinity of the oligonucleotide for a target nucleic acidsequence, such as poly-(L-lysine). Further still, intercalating agents,such as ellipticine, and alkylating agents or metal complexes may beattached to sense or antisense oligonucleotides to modify bindingspecificities of the antisense or sense oligonucleotide for the targetnucleotide sequence.

[0419] Antisense or sense oligonucleotides may be introduced into a cellcontaining the target nucleic acid sequence by any gene transfer method,including, for example, CaPO₄-mediated DNA transfection,electroporation, or by using gene transfer vectors such as Epstein-Barrvirus. In a preferred procedure, an antisense or sense oligonucleotideis inserted into a suitable retroviral vector. A cell containing thetarget nucleic acid sequence is contacted with the recombinantretroviral vector, either in vivo or ex vivo. Suitable retroviralvectors include, but are not limited to, those derived from the murineretrovirus M-MuLV, N2 (a retrovirus derived from M-MuLV), or the doublecopy vectors designated DCT5A, DCT5B and DCT5C (see WO 90/13641).

[0420] Sense or antisense oligonucleotides also may be introduced into acell containing the target nucleotide sequence by formation of aconjugate with a ligand binding molecule, as described in WO 91/04753.Suitable ligand binding molecules include, but are not limited to, cellsurface receptors, growth factors, other cytokines, or other ligandsthat bind to cell surface receptors. Preferably, conjugation of theligand binding molecule does not substantially interfere with theability of the ligand binding molecule to bind to its correspondingmolecule or receptor, or block entry of the sense or antisenseoligonucleotide or its conjugated version into the cell.

[0421] Alternatively, a sense or an antisense oligonucleotide may beintroduced into a cell containing the target nucleic acid sequence byformation of an oligonucleotide-lipid complex, as described in WO90/10448. The sense or antisense oligonucleotide-lipid complex ispreferably dissociated within the cell by an endogenous lipase.

[0422] Antisense or sense RNA or DNA molecules are generally at leastabout 5 bases in length, about 10 bases in length, about 15 bases inlength, about 20 bases in length, about 25 bases in length, about 30bases in length, about 35 bases in length, about 40 bases in length,about 45 bases in length, about 50 bases in length, about 55 bases inlength, about 60 bases in length, about 65 bases in length, about 70bases in length, about 75 bases in length, about 80 bases in length,about 85 bases in length, about 90 bases in length, about 95 bases inlength, about 100 bases in length, or more.

[0423] The probes may also be employed in PCR techniques to generate apool of sequences for identification of closely related PRO codingsequences.

[0424] Nucleotide sequences encoding a PRO can also be used to constructhybridization probes for mapping the gene which encodes that PRO and forthe genetic analysis of individuals with genetic disorders. Thenucleotide sequences provided herein may be mapped to a chromosome andspecific regions of a chromosome using known techniques, such as in situhybridization, linkage analysis against known chromosomal markers, andhybridization screening with libraries.

[0425] When the coding sequences for PRO encode a protein which binds toanother protein (example, where the PRO is a receptor), the PRO can beused in assays to identify the other proteins or molecules involved inthe binding interaction. By such methods, inhibitors of thereceptor/ligand binding interaction can be identified. Proteins involvedin such binding interactions can also be used to screen for peptide orsmall molecule inhibitors or agonists of the binding interaction. Also,the receptor PRO can be used to isolate correlative ligand(s). Screeningassays can be designed to find lead compounds that mimic the biologicalactivity of a native PRO or a receptor for PRO. Such screening assayswill include assays amenable to high-throughput screening of chemicallibraries, making them particularly suitable for identifying smallmolecule drug candidates. Small molecules contemplated include syntheticorganic or inorganic compounds. The assays can be performed in a varietyof formats, including protein-protein binding assays, biochemicalscreening assays, immunoassays and cell based assays, which are wellcharacterized in the art.

[0426] Nucleic acids which encode PRO or its modified forms can also beused to generate either transgenic animals or “knock out” animals which,in turn, are useful in the development and screening of therapeuticallyuseful reagents. A transgenic animal (e.g., a mouse or rat) is an animalhaving cells that contain a transgene, which transgene was introducedinto the animal or an ancestor of the animal at a prenatal, e.g., anembryonic stage. A transgene is a DNA which is integrated into thegenome of a cell from which a transgenic animal develops. In oneembodiment, cDNA encoding PRO can be used to clone genomic DNA encodingPRO in accordance with established techniques and the genomic sequencesused to generate transgenic animals that contain cells which express DNAencoding PRO. Methods for generating transgenic animals, particularlyanimals such as mice or rats, have become conventional in the art andare described, for example, in U.S. Pat. Nos. 4,736,866 and 4,870,009.Typically, particular cells would be targeted for PRO transgeneincorporation with tissue-specific enhancers. Transgenic animals thatinclude a copy of a transgene encoding PRO introduced into the germ lineof the animal at an embryonic stage can be used to examine the effect ofincreased expression of DNA encoding PRO. Such animals can be used astester animals for reagents thought to confer protection from, forexample, pathological conditions associated with its overexpression. Inaccordance with this facet of the invention, an animal is treated withthe reagent and a reduced incidence of the pathological condition,compared to untreated animals bearing the transgene, would indicate apotential therapeutic intervention for the pathological condition.

[0427] Alternatively, non-human homologues of PRO can be used toconstruct a PRO “knock out” animal which has a defective or altered geneencoding PRO as a result of homologous recombination between theendogenous gene encoding PRO and altered genomic DNA encoding PROintroduced into an embryonic stem cell of the animal. For example, cDNAencoding PRO can be used to clone genomic DNA encoding PRO in accordancewith established techniques. A portion of the genomic DNA encoding PROcan be deleted or replaced with another gene, such as a gene encoding aselectable marker which can be used to monitor integration. Typically,several kilobases of unaltered flanking DNA (both at the 5′ and 3′ ends)are included in the vector [see e.g., Thomas and Capecchi, Cell, 51:503(1987) for a description of homologous recombination vectors]. Thevector is introduced into an embryonic stem cell line (e.g., byelectroporation) and cells in which the introduced DNA has homologouslyrecombined with the endogenous DNA are selected [see e.g., Li et al.,Cell, 69:915 (1992)]. The selected cells are then injected into ablastocyst of an animal (e.g., a mouse or rat) to form aggregationchimeras [see e.g., Bradley, in Teratocarcinomas and Embryonic StemCells: A Practical Approach, E. J. Robertson, ed. (IRL, Oxford, 1987),pp. 113-152]. A chimeric embryo can then be implanted into a suitablepseudopregnant female foster animal and the embryo brought to term tocreate a “knock out” animal. Progeny harboring the homologouslyrecombined DNA in their germ cells can be identified by standardtechniques and used to breed animals in which all cells of the animalcontain the homologously recombined DNA. Knockout animals can becharacterized for instance, for their ability to defend against certainpathological conditions and for their development of pathologicalconditions due to absence of the PRO polypeptide.

[0428] Nucleic acid encoding the PRO polypeptides may also be used ingene therapy. In gene therapy applications, genes are introduced intocells in order to achieve in vivo synthesis of a therapeuticallyeffective genetic product, for example for replacement of a defectivegene. “Gene therapy” includes both conventional gene therapy where alasting effect is achieved by a single treatment, and the administrationof gene therapeutic agents, which involves the one time or repeatedadministration of a therapeutically effective DNA or mRNA. AntisenseRNAs and DNAs can be used as therapeutic agents for blocking theexpression of certain genes in vivo. It has already been shown thatshort antisense oligonucleotides can be imported into cells where theyact as inhibitors, despite their low intracellular concentrations causedby their restricted uptake by the cell membrane. (Zamecnik et al., Proc.Natl. Acad. Sci. USA 83:4143-4146 [1986]). The oligonucleotides can bemodified to enhance their uptake, e.g. by substituting their negativelycharged pliosphodiester groups by uncharged groups.

[0429] There are a variety of techniques available for introducingnucleic acids into viable cells. The techniques vary depending uponwhether the nucleic acid is transferred into cultured cells in vitro, orin vivo in the cells of the intended host. Techniques suitable for thetransfer of nucleic acid into mammalian cells in vitro include the useof liposomes, electroporation, microinjection, cell fusion,DEAE-dextran, the calcium phosphate precipitation method, etc. Thecurrently preferred in vivo gene transfer techniques includetransfection with viral (typically retroviral) vectors and viral coatprotein-liposome mediated transfection (Dzau et al., Trends inBiotechnology 11, 205-210 [1993]). In some situations it is desirable toprovide the nucleic acid source with an agent that targets the targetcells, such as an antibody specific for a cell surface membrane proteinor the target cell, a ligand for a receptor on the target cell, etc.Where liposomes are employed, proteins which bind to a cell surfacemembrane protein associated with endocytosis may be used for targetingand/or to facilitate uptake, e.g. capsid proteins or fragments thereoftropic for a particular cell type, antibodies for proteins which undergointernalization in cycling, proteins that target intracellularlocalization and enhance intracellular half-life. The technique ofreceptor-mediated endocytosis is described, for example, by Wu et al.,J. Biol. Chem. 262, 4429-4432 (1987); and Wagner et al., Proc. Natl.Acad. Sci. USA 87, 3410-3414 (1990). For review of gene marking and genetherapy protocols see Anderson et al., Science 256, 808-813 (1992).

[0430] The PRO polypeptides described herein may also be employed asmolecular weight markers for protein electrophoresis purposes and theisolated nucleic acid sequences may be used for recombinantly expressingthose markers.

[0431] The nucleic acid molecules encoding the PRO polypeptides orfragments thereof described herein are useful for chromosomeidentification. In this regard, there exists an ongoing need to identifynew chromosome markers, since relatively few chromosome markingreagents, based upon actual sequence data are presently available. EachPRO nucleic acid molecule of the present invention can be used as achromosome marker.

[0432] The PRO polypeptides and nucleic acid molecules of the presentinvention may also be used for tissue typing, wherein the PROpolypeptides of the present invention may be differentially expressed inone tissue as compared to another. PRO nucleic acid molecules will finduse for generating probes for PCR, Northern analysis, Southern analysisand Western analysis.

[0433] The PRO polypeptides described herein may also be employed astherapeutic agents. The PRO polypeptides of the present invention can beformulated according to known methods to prepare pharmaceutically usefulcompositions, whereby the PRO product hereof is combined in admixturewith a pharmaceutically acceptable carrier vehicle. Therapeuticformulations are prepared for storage by mixing the active ingredienthaving the desired degree of purity with optional physiologicallyacceptable carriers, excipients or stabilizers (Remington'sPharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the formof lyophilized formulations or aqueous solutions. Acceptable carriers,excipients or stabilizers are nontoxic to recipients at the dosages andconcentrations employed, and include buffers such as phosphate, citrateand other organic acids; antioxidants including ascorbic acid; lowmolecular weight (less than about 10 residues) polypeptides; proteins,such as serum albumin, gelatin or immunoglobulins; hydrophilic polymerssuch as polyvinylpyrrolidone, amino acids such as glycine, glutamine,asparagine, arginine or lysine; monosaccharides, disaccharides and othercarbohydrates including glucose, mannose, or dextrins; chelating agentssuch as EDTA; sugar alcohols such as mannitol or sorbitol; salt-formingcounterions such as sodium; and/or nonionic surfactants such as TWEEN™,PLURONICS™ or PEG.

[0434] The formulations to be used for in vivo administration must besterile. This is readily accomplished by filtration through sterilefiltration membranes, prior to or following lyophilization andreconstitution.

[0435] Therapeutic compositions herein generally are placed into acontainer having a sterile access port, for example, an intravenoussolution bag or vial having a stopper pierceable by a hypodermicinjection needle.

[0436] The route of administration is in accord with known methods, e.g.injection or infusion by intravenous, intraperitoneal, intracerebral,intramuscular, intraocular, intraarterial or intralesional routes,topical administration, or by sustained release systems.

[0437] Dosages and desired drug concentrations of pharmaceuticalcompositions of the present invention may vary depending on theparticular use envisioned. The determination of the appropriate dosageor route of administration is well within the skill of an ordinaryphysician. Animal experiments provide reliable guidance for thedetermination of effective doses for human therapy. Interspecies scalingof effective doses can be performed following the principles laid downby Mordenti, J. and Chappell, W. “The use of interspecies scaling intoxicokinetics” In Toxicokinetics and New Drug Development, Yacobi etal., Eds., Pergamon Press, New York 1989, pp. 42-96.

[0438] When in vivo administration of a PRO polypeptide or agonist orantagonist thereof is employed, normal dosage amounts may vary fromabout 10 ng/kg to up to 100 mg/kg of mammal body weight or more per day,preferably about 1 μg/kg/day to 10 mg/kg/day, depending upon the routeof administration. Guidance as to particular dosages and methods ofdelivery is provided in the literature; see, for example, U.S. Pat. Nos.4,657,760; 5,206,344; or 5,225,212. It is anticipated that differentformulations will be effective for different treatment compounds anddifferent disorders, that administration targeting one organ or tissue,for example, may necessitate delivery in a manner different from that toanother organ or tissue.

[0439] Where sustained-release administration of a PRO polypeptide isdesired in a formulation with release characteristics suitable for thetreatment of any disease or disorder requiring administration of the PROpolypeptide, microencapsulation of the PRO polypeptide is contemplated.Microencapsulation of recombinant proteins for sustained release hasbeen successfully performed with human growth hormone (rhGH),interferon-(rhlFN- ), interleukin-2, and MN rgp 120. Johnson et al.,Nat. Med., 2:795-799 (1996); Yasuda, Biomed. Ther., 27:1221-1223 (1993);Hora et al., Bio/Technology. 8:755-758(1990); Cleland, “Design andProduction of Single Immunization Vaccines Using PolylactidePolyglycolide Microsphere Systems,” in Vaccine Design: The Subunit andAdjuvant Approach, Powell and Newman, eds, (Plenum Press: New York,1995), pp. 439-462; WO 97/03692, WO 96/40072, WO 96/07399; and U.S. Pat.No. 5,654,010.

[0440] The sustained-release formulations of these proteins weredeveloped using poly-lactic-coglycolic acid (PLGA) polymer due to itsbiocompatibility and wide range of biodegradable properties. Thedegradation products of PLGA, lactic and glycolic acids, can be clearedquickly within the human body. Moreover, the degradability of thispolymer can be adjusted from months to years depending on its molecularweight and composition. Lewis, “Controlled release of bioactive agentsfrom lactide/glycolide polymer,” in: M. Chasin and R. Langer (Eds.),Biodegradable Polymers as Drug Delivery Systems (Marcel Dekker: NewYork, 1990), pp. 1-41.

[0441] This invention encompasses methods of screening compounds toidentify those that mimic the PRO polypeptide (agonists) or prevent theeffect of the PRO polypeptide (antagonists). Screening assays forantagonist drug candidates are designed to identify compounds that bindor complex with the PRO polypeptides encoded by the genes identifiedherein, or otherwise interfere with the interaction of the encodedpolypeptides with other cellular proteins. Such screening assays willinclude assays amenable to high-throughput screening of chemicallibraries, making them particularly suitable for identifying smallmolecule drug candidates.

[0442] The assays can be performed in a variety of formats, includingprotein-protein binding assays, biochemical screening assays,immunoassays, and cell-based assays, which are well characterized in theart.

[0443] All assays for antagonists are common in that they call forcontacting the drug candidate with a PRO polypeptide encoded by anucleic acid identified herein under conditions and for a timesufficient to allow these two components to interact.

[0444] In binding assays, the interaction is binding and the complexformed can be isolated or detected in the reaction mixture. In aparticular embodiment, the PRO polypeptide encoded by the geneidentified herein or the drug candidate is immobilized on a solid phase,e.g., on a microtiter plate, by covalent or non-covalent attachments.Non-covalent attachment generally is accomplished by coating the solidsurface with a solution of the PRO polypeptide and drying.Alternatively, an immobilized antibody, e.g., a monoclonal antibody,specific for the PRO polypeptide to be immobilized can be used to anchorit to a solid surface. The assay is performed by adding thenon-immobilized component, which may be labeled by a detectable label,to the immobilized component, e.g., the coated surface containing theanchored component. When the reaction is complete, the non-reactedcomponents are removed, e.g., by washing, and complexes anchored on thesolid surface are detected. When the originally non-immobilizedcomponent carries a detectable label, the detection of label immobilizedon the surface indicates that complexing occurred. Where the originallynon-immobilized component does not carry a label, complexing can bedetected, for example, by using a labeled antibody specifically bindingthe immobilized complex.

[0445] If the candidate compound interacts with but does not bind to aparticular PRO polypeptide encoded by a gene identified herein, itsinteraction with that polypeptide can be assayed by methods well knownfor detecting protein-protein interactions. Such assays includetraditional approaches, such as, e.g., cross-linking,co-immunoprecipitation, and co-purification through gradients orchromatographic columns. In addition, protein-protein interactions canbe monitored by using a yeast-based genetic system described by Fieldsand co-workers (Fields and Song, Nature (London), 340:245-246 (1989);Chien et al., Proc. Natl. Acad. Sci. USA, 88:9578-9582 (1991)) asdisclosed by Chevray and Nathans, Proc. Natl. Acad. Sci. USA, 89:5789-5793(1991). Many transcriptional activators, such as yeast GAL4,consist of two physically discrete modular domains, one acting as theDNA-binding domain, the other one functioning as thetranscription-activation domain. The yeast expression system describedin the foregoing publications (generally referred to as the “two-hybridsystem”) takes advantage of this property, and employs two hybridproteins, one in which the target protein is fused to the DNA-bindingdomain of GAL4, and another, in which candidate activating proteins arefused to the activation domain. The expression of a GAL1-lacZ reportergene under control of a GAL4-activated promoter depends onreconstitution of GAL4 activity via protein-protein interaction.Colonies containing interacting polypeptides are detected with achromogenic substrate for β-galactosidase. A complete kit (MATCHMAKER™)for identifying protein-protein interactions between two specificproteins using the two-hybrid technique is commercially available fromClontech. This system can also be extended to map protein domainsinvolved in specific protein interactions as well as to pinpoint aminoacid residues that are crucial for these interactions.

[0446] Compounds that interfere with the interaction of a gene encodinga PRO polypeptide identified herein and other intra- or extracellularcomponents can be tested as follows: usually a reaction mixture isprepared containing the product of the gene and the intra- orextracellular component under conditions and for a time allowing for theinteraction and binding of the two products. To test the ability of acandidate compound to inhibit binding, the reaction is run in theabsence and in the presence of the test compound. In addition, a placebomay be added to a third reaction mixture, to serve as positive control.The binding (complex formation) between the test compound and the intra-or extracellular component present in the mixture is monitored asdescribed hereinabove. The formation of a complex in the controlreaction(s) but not in the reaction mixture containing the test compoundindicates that the test compound interferes with the interaction of thetest compound and its reaction partner.

[0447] To assay for antagonists, the PRO polypeptide may be added to acell along with the compound to be screened for a particular activityand the ability of the compound to inhibit the activity of interest inthe presence of the PRO polypeptide indicates that the compound is anantagonist to the PRO polypeptide. Alternatively, antagonists may bedetected by combining the PRO polypeptide and a potential antagonistwith membrane-bound PRO polypeptide receptors or recombinant receptorsunder appropriate conditions for a competitive inhibition assay. The PROpolypeptide can be labeled, such as by radioactivity, such that thenumber of PRO polypeptide molecules bound to the receptor can be used todeternine the effectiveness of the potential antagonist. The geneencoding the receptor can be identified by numerous methods known tothose of skill in the art, for example, ligand panning and FACS sorting.Coligan et al., Current Protocols in Immun., 1(2): Chapter 5 (1991).Preferably, expression cloning is employed wherein polyadenylated RNA isprepared from a cell responsive to the PRO polypeptide and a cDNAlibrary created from this RNA is divided into pools and used totransfect COS cells or other cells that are not responsive to the PROpolypeptide. Transfected cells that are grown on glass slides areexposed to labeled PRO polypeptide. The PRO polypeptide can be labeledby a variety of means including iodination or inclusion of a recognitionsite for a site-specific protein kinase. Following fixation andincubation, the slides are subjected to autoradiographic analysis.Positive pools are identified and sub-pools are prepared andre-transfected using an interactive sub-pooling and re-screeningprocess, eventually yielding a single clone that encodes the putativereceptor.

[0448] As an alternative approach for receptor identification, labeledPRO polypeptide can be photoaffinity-linked with cell membrane orextract preparations that express the receptor molecule. Cross-linkedmaterial is resolved by PAGE and exposed to X-ray film. The labeledcomplex containing the receptor can be excised, resolved into peptidefragments, and subjected to protein micro-sequencing. The amino acidsequence obtained from micro-sequencing would be used to design a set ofdegenerate oligonucleotide probes to screen a cDNA library to identifythe gene encoding the putative receptor.

[0449] In another assay for antagonists, mammalian cells or a membranepreparation expressing the receptor would be incubated with labeled PROpolypeptide in the presence of the candidate compound. The ability ofthe compound to enhance or block this interaction could then bemeasured.

[0450] More specific examples of potential antagonists include anoligonucleotide that binds to the fusions of immunoglobulin with PROpolypeptide, and, in particular, antibodies including, withoutlimitation, poly-and monoclonal antibodies and antibody fragments,single-chain antibodies, anti-idiotypic antibodies, and chimeric orhumanized versions of such antibodies or fragments, as well as humanantibodies and antibody fragments. Alternatively, a potential antagonistmay be a closely related protein, for example, a mutated form of the PROpolypeptide that recognizes the receptor but imparts no effect, therebycompetitively inhibiting the action of the PRO polypeptide.

[0451] Another potential PRO polypeptide antagonist is an antisense RNAor DNA construct prepared using antisense technology, where, e.g., anantisense RNA or DNA molecule acts to block directly the translation ofmRNA by hybridizing to targeted mRNA and preventing protein translation.Antisense technology can be used to control gene expression throughtriple-helix formation or antisense DNA or RNA, both of which methodsare based on binding of a polynucleotide to DNA or RNA. For example, the5′ coding portion of the polynucleotide sequence, which encodes themature PRO polypeptides herein, is used to design an antisense RNAoligonucleotide of from about 10 to 40 base pairs in length. A DNAoligonucleotide is designed to be complementary to a region of the geneinvolved in transcription (triple helix-see Lee et al., Nucl. AcidsRes., 6:3073 (1979); Cooney et al., Science, 241: 456 (1988); Dervan etal., Science, 251:1360 (1991)), thereby preventing transcription and theproduction of the PRO polypeptide. The antisense RNA oligonucleotidehybridizes to the mRNA in vivo and blocks translation of the mRNAmolecule into the PRO polypeptide (antisense-Okano, Neurochem., 56:560(1991); Oligodeoxynucleotides as Antisense Inhibitors of Gene Expression(CRC Press: Boca Raton, Fla., 1988). The oligonucleotides describedabove can also be delivered to cells such that the antisense RNA or DNAmay be expressed in vivo to inhibit production of the PRO polypeptide.When antisense DNA is used, oligodeoxyribonucleotides derived from thetranslation-initiation site, e.g., between about −10 and +10 positionsof the target gene nucleotide sequence, are preferred.

[0452] Potential antagonists include small molecules that bind to theactive site, the receptor binding site, or growth factor or otherrelevant binding site of the PRO polypeptide, thereby blocking thenormal biological activity of the PRO polypeptide. Examples of smallmolecules include, but are not limited to, small peptides orpeptide-like molecules, preferably soluble peptides, and syntheticnon-peptidyl organic or inorganic compounds.

[0453] Ribozymes are enzymatic RNA molecules capable of catalyzing thespecific cleavage of RNA. Ribozymes act by sequence-specifichybridization to the complementary target RNA, followed byendonucleolytic cleavage. Specific ribozyme cleavage sites within apotential RNA target can be identified by known techniques. For furtherdetails see, e.g., Rossi, Current Biology, 4:469-471 (1994), and PCTpublication No. WO 97/33551 (published Sep. 18, 1997).

[0454] Nucleic acid molecules in triple-helix formation used to inhibittranscription should be single-stranded and composed ofdeoxynucleotides. The base composition of these oligonucleotides isdesigned such that it promotes triple-helix formation via Hoogsteenbase-pairing rules, which generally require sizeable stretches ofpurines or pyrimidines on one strand of a duplex. For further detailssee, e.g., PCT publication No. WO 97/33551, supra.

[0455] These small molecules can be identified by any one or more of thescreening assays discussed hereinabove and/or by any other screeningtechniques well known for those skilled in the art.

[0456] Uses of the herein disclosed molecules may also be based upon thepositive functional assay hits disclosed and described below. Methodsbased upon those assay hits are also encompassed by the presentinvention.

[0457] F. Anti-PRO Antibodies

[0458] The present invention further provides anti-PRO antibodies.Exemplary antibodies include polyclonal, monoclonal, humanized,bispecific, and heteroconjugate antibodies.

[0459] 1. Polyclonal Antibodies

[0460] The anti-PRO antibodies may comprise polyclonal antibodies.Methods of preparing polyclonal antibodies are known to the skilledartisan. Polyclonal antibodies can be raised in a mammal, for example,by one or more injections of an immunizing agent and, if desired, anadjuvant. Typically, the immunizing agent and/or adjuvant will beinjected in the mammal by multiple subcutaneous or intraperitonealinjections. The immunizing agent may include the PRO polypeptide or afusion protein thereof. It may be useful to conjugate the immunizingagent to a protein known to be immunogenic in the mammal beingimmunized. Examples of such immunogenic proteins include but are notlimited to keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. Examples of adjuvants which may beemployed include Freund's complete adjuvant and MPL-TDM adjuvant(monophosphoryl Lipid A, synthetic trehalose dicorynomycolate). Theimmunization protocol may be selected by one skilled in the art withoutundue experimentation.

[0461] 2. Monoclonal Antibodies

[0462] The anti-PRO antibodies may, alternatively, be monoclonalantibodies. Monoclonal antibodies may be prepared using hybridomamethods, such as those described by Kohler and Milstein, Nature, 256:495(1975). In a hybridoma method, a mouse, hamster, or other appropriatehost animal, is typically immunized with an immunizing agent to elicitlymphocytes that produce or are capable of producing antibodies thatwill specifically bind to the immunizing agent. Alternatively, thelymphocytes may be immunized in vitro.

[0463] The immunizing agent will typically include the PRO polypeptideor a fusion protein thereof. Generally, either peripheral bloodlymphocytes (“PBLs”) are used if cells of human origin are desired, orspleen cells or lymph node cells are used if non-human mammalian sourcesare desired. The lymphocytes are then fused with an immortalized cellline using a suitable fusing agent, such as polyethylene glycol, to forma hybridoma cell [Goding, Monoclonal Antibodies: Principles andPractice, Academic Press, (1986) pp. 59-103]. Immortalized cell linesare usually transformed mammalian cells, particularly mycloma cells ofrodent, bovine and human origin. Usually, rat or mouse myeloma celllines are employed. The hybridoma cells may be cultured in a suitableculture medium that preferably contains one or more substances thatinhibit the growth or survival of the unfused, immortalized cells. Forexample, if the parental cells lack the enzyme hypoxanthine guaninephosphoribosyl transferase (HGPRT or HPRT), the culture medium for thehybridomas typically will include hypoxanthine, aminopterin, andthymidine (“HAT medium”), which substances prevent the growth ofHGPRT-deficient cells.

[0464] Preferred immortalized cell lines are those that fuseefficiently, support stable high level expression of antibody by theselected antibody-producing cells, and are sensitive to a medium such asHAT medium. More preferred immortalized cell lines are murine myelomalines, which can be obtained, for instance, from the Salk Institute CellDistribution Center, San Diego, California and the American Type CultureCollection, Manassas, Virginia. Human myeloma and mouse-humanheteromyeloma cell lines also have been described for the production ofhuman monoclonal antibodies [Kozbor, J. Immunol., 133:3001 (1984);Brodeur et al., Monoclonal Antibody Production Techniques andApplications, Marcel Dekker, Inc., New York, (1987) pp. 51-63].

[0465] The culture medium in which the hybridoma cells are cultured canthen be assayed for the presence of monoclonal antibodies directedagainst PRO. Preferably, the binding specificity of monoclonalantibodies produced by the hybridoma cells is determined byimmunoprecipitation or by an in vitro binding assay, such asradioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA).Such techniques and assays are known in the art. The binding affinity ofthe monoclonal antibody can, for example, be determined by the Scatchardanalysis of Munson and Pollard, Anal. Biochem., 107:220 (1980).

[0466] After the desired hybridoma cells are identified, the clones maybe subcloned by limiting dilution procedures and grown by standardmethods [Goding, supra]. Suitable culture media for this purposeinclude, for example, Dulbecco's Modified Eagle's Medium and RPMI-1640medium. Alternatively, the hybridoma cells may be grown in vivo asascites in a mammal.

[0467] The monoclonal antibodies secreted by the subclones may beisolated or purified from the culture medium or ascites fluid byconventional immunoglobulin purification procedures such as, forexample, protein A-Sepharose, hydroxylapatite chromatography, gelelectrophoresis, dialysis, or affinity chromatography.

[0468] The monoclonal antibodies may also be made by recombinant DNAmethods, such as those described in U.S. Pat. No. 4,816,567. DNAencoding the monoclonal antibodies of the invention can be readilyisolated and sequenced using conventional procedures (e.g., by usingoligonucleotide probes that are capable of binding specifically to genesencoding the heavy and light chains of murine antibodies). The hybridomacells of the invention serve as a preferred source of such DNA. Onceisolated, the DNA may be placed into expression vectors, which are thentransfected into host cells such as simian COS cells, Chinese hamsterovary (CHO) cells, or myeloma cells that do not otherwise produceimmunoglobulin protein, to obtain the synthesis of monoclonal,antibodiesin the recombinant host cells. The DNA also may be modified, forexample, by substituting the coding sequence for human heavy and lightchain constant domains in place of the homologous murine sequences [U.S.Pat. No. 4,816,567; Morrison et al., supra] or by covalently joining tothe immunoglobulin coding sequence all or part of the coding sequencefor a non-immunoglobulin polypeptide. Such a non-immunoglobulinpolypeptide can be substituted for the constant domains of an antibodyof the invention, or can be substituted for the variable domains of oneantigen-combining site of an antibody of the invention to create achimeric bivalent antibody.

[0469] The antibodies may be monovalent antibodies. Methods forpreparing monovalent antibodies are well known in the art. For example,one method involves recombinant expression of immunoglobulin light chainand modified heavy chain. The heavy chain is truncated generally at anypoint in the Fc region so as to prevent heavy chain crosslinking.Alternatively, the relevant cysteine residues are substituted withanother amino acid residue or are deleted so as to prevent crosslinking.

[0470] In vitro methods are also suitable for preparing monovalentantibodies. Digestion of antibodies to produce fragments thereof,particularly, Fab fragments, can be accomplished using routinetechniques known in the art.

[0471] 3. Human and Humanized Antibodies

[0472] The anti-PRO antibodies of the invention may further comprisehumanized antibodies or human antibodies. Humanized forms of non-human(e.g., murine) antibodies are chimeric immunoglobulins, immunoglobulinchains or fragments thereof (such as Fv, Fab, Fab′, F(ab′)₂ or otherantigen-binding subsequences of antibodies) which contain minimalsequence derived from non-human immunoglobulin. Humanized antibodiesinclude human immunoglobulins (recipient antibody) in which residuesfrom a complementary determining region (CDR) of the recipient arereplaced by residues from a CDR of a non-human species (donor antibody)such as mouse, rat or rabbit having the desired specificity, affinityand capacity. In some instances, Fv framework residues of the humanimmunoglobulin are replaced by corresponding non-human residues.Humanized antibodies may also comprise residues which are found neitherin the recipient antibody nor in the imported CDR or frameworksequences. In general, the humanized antibody will comprisesubstantially all of at least one, and typically two, variable domains,in which all or substantially all of the CDR regions correspond to thoseof a non-human immunoglobulin and all or substantially all of the FRregions are those of a human immunoglobulin consensus sequence. Thehumanized antibody optimally also will comprise at least a portion of animmunoglobulin constant region (Fc), typically that of a humanimmunoglobulin [Jones et al., Nature, 321:522-525 (1986); Riechmann etal., Nature, 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol.,2:593-596 (1992)].

[0473] Methods for humanizing non-human antibodies are well known in theart. Generally, a humanized antibody has one or more amino acid residuesintroduced into it from a source which is non-human. These non-humanamino acid residues are often referred to as “import” residues, whichare typically taken from an “import” variable domain. Humanization canbe essentially performed following the method of Winter and co-workers[Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature,332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536 (1988)], bysubstituting rodent CDRs or CDR sequences for the correspondingsequences of a human antibody. Accordingly, such “humanized” antibodiesare chimeric antibodies (U.S. Pat. No. 4,816,567), wherein substantiallyless than an intact human variable domain has been substituted by thecorresponding sequence from a non-human species. In practice, humanizedantibodies are typically human antibodies in which some CDR residues andpossibly some FR residues are substituted by residues from analogoussites in rodent antibodies.

[0474] Human antibodies can also be produced using various techniquesknown in the art, including phage display libraries [Hoogenboom andWinter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol.,222:581 (1991)]. The techniques of Cole et al. and Boerner et al. arealso available for the preparation of human monoclonal antibodies (Coleet al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77(1985) and Boerner et al., J. Immunol., 147(1):86-95 (1991)]. Similarly,human antibodies can be made by introducing of human immunoglobulin lociinto transgenic animals, e.g., mice in which the endogenousimmunoglobulin genes have been partially or completely inactivated. Uponchallenge, human antibody production is observed, which closelyresembles that seen in humans in all respects, including generearrangement, assembly, and antibody repertoire. This approach isdescribed, for example, in U.S. Pat. Nos. 5,545,807; 5,545,806;5,569,825; 5,625,126; 5,633,425; 5,661,016, and in the followingscientific publications: Marks et al., Bio/Technology 10, 779-783(1992); Lonberg et al., Nature 368 856-859 (1994); Morrison, Nature 368,812-13 (1994); Fishwild et al., Nature Biotechnology 14, 845-51 (1996);Neuberger, Nature Biotechnology 14, 826 (1996); Lonberg and Huszar,Intern. Rev. Immunol. 13 65-93 (1995).

[0475] The antibodies may also be affinity matured using known selectionand/or mutagenesis methods as described above. Preferred affinitymatured antibodies have an affinity which is five times, more preferably10 times, even more preferably 20 or 30 times greater than the startingantibody (generally murine, humanized or human) from which the maturedantibody is prepared.

[0476] 4. Bispecific Antibodies

[0477] Bispecific antibodies are monoclonal, preferably human orhumanized, antibodies that have binding specificities for at least twodifferent antigens. In the present case, one of the bindingspecificities is for the PRO, the other one is for any other antigen,and preferably for a cell-surface protein or receptor or receptorsubunit.

[0478] Methods for making bispecific antibodies are known in the art.Traditionally, the recombinant production of bispecific antibodies isbased on the co-expression of two immunoglobulin heavy-chain/light-chainpairs, where the two heavy chains have different specificities [Milsteinand Cuello, Nature, 305:537-539 (1983)]. Because of the randomassortment of immunoglobulin heavy and light chains, these hybridomas(quadromas) produce a potential mixture of ten different antibodymolecules, of which only one has the correct bispecific structure. Thepurification of the correct molecule is usually accomplished by affinitychromatography steps. Similar procedures are disclosed in WO 93/08829,published May 13, 1993, and in Traunecker et al., EMBO J., 10:3655-3659(1991).

[0479] Antibody variable domains with the desired binding specificities(antibody-antigen combining sites) can be fused to immunoglobulinconstant domain sequences. The fusion preferably is with animmunoglobulin heavy-chain constant domain, comprising at least part ofthe hinge, CH2, and CH3 regions. It is preferred to have the firstheavy-chain constant region (CH 1) containing the site necessary forlight-chain binding present in at least one of the fusions. DNAsencoding the immunoglobulin heavy-chain fusions and, if desired, theimmunoglobulin light chain, are inserted into separate expressionvectors, and are co-transfected into a suitable host organism. Forfurther details of generating bispecific antibodies see, for example,Suresh et al., Methods in Enzymology, 121:210 (1986).

[0480] According to another approach described in WO 96/27011, theinterface between a pair of antibody molecules can be engineered tomaximize the percentage of heterodimers which are recovered fromrecombinant cell culture. The preferred interface comprises at least apart of the CH3 region of an antibody constant domain. In this method,one or more small amino acid side chains from the interface of the firstantibody molecule are replaced with larger side chains (e.g. tyrosine ortryptophan). Compensatory “cavities” of identical or similar size to thelarge side chain(s) are created on the interface of the second antibodymolecule by replacing large amino acid side chains with smaller ones(e.g. alanine or threonine). This provides a mechanism for increasingthe yield of the heterodimer over other unwanted end-products such ashomodimers.

[0481] Bispecific antibodies can be prepared as full length antibodiesor antibody fragments (e.g. F(ab′)₂ bispecific antibodies). Techniquesfor generating bispecific antibodies from antibody fragments have beendescribed in the literature. For example, bispecific antibodies can beprepared can be prepared using chemical linkage. Brennan et al., Science229:81 (1985) describe a procedure wherein intact antibodies areproteolytically cleaved to generate F(ab′)₂ fragments. These fragmentsare reduced in the presence of the dithiol complexing agent sodiumarsenite to stabilize vicinal dithiols and prevent intermoleculardisulfide formation. The Fab′ fragments generated are then converted tothionitrobenzoate (TNB) derivatives. One of the Fab′-TNB derivatives isthen reconverted to the Fab′-thiol by reduction with mercaptoethylamineand is mixed with an equimolar amount of the other Fab′-TNB derivativeto form the bispecific antibody. The bispecific antibodies produced canbe used as agents for the selective immobilization of enzymes.

[0482] Fab′ fragments may be directly recovered from E. coli andchemically coupled to form bispecific antibodies. Shalaby et al., J.Exp. Med. 175:217-225 (1992) describe the production of a fullyhumanized bispecific antibody F(ab′)₂ molecule. Each Fab′ fragment wasseparately secreted from E. coli and subjected to directed chemicalcoupling in vitro to form the bispecific antibody. The bispecificantibody thus formed was able to bind to cells overexpressing the ErbB2receptor and normal human T cells, as well as trigger the lytic activityof human cytotoxic lymphocytes against human breast tumor targets.

[0483] Various technique for making and isolating bispecific antibodyfragments directly from recombinant cell culture have also beendescribed. For example, bispecific antibodies have been produced usingleucine zippers. Kostelny et al., J. Immunol. 148(5):1547-1553 (1992).The leucine zipper peptides from the Fos and Jun proteins were linked tothe Fab′ portions of two different antibodies by gene fusion. Theantibody homodimers were reduced at the hinge region to form monomersand then re-oxidized to form the antibody heterodimers. This method canalso be utilized for the production of antibody homodimers. The“diabody” technology described by Hollinger et al., Proc. Natl. Acad.Sci. USA 90:6444-6448 (1993) has provided an alternative mechanism formaking bispecific antibody fragments. The fragments comprise aheavy-chain variable domain (V_(H)) connected to a light-chain variabledomain (V_(L)) by a linker which is too short to allow pairing betweenthe two domains on the same chain. Accordingly, the V_(H) and V_(L)domains of one fragment are forced to pair with the complementary V_(L)and V_(H) domains of another fragment, thereby forming twoantigen-binding sites. Another strategy for making bispecific antibodyfragments by the use of single-chain Fv (sFv) dimers has also beenreported. See, Gruber et al., J. Immunol. 152:5368 (1994). Antibodieswith more than two valencies are contemplated. For example, trispecificantibodies can be prepared. Tutt et al., J. Immunol. 147:60 (1991).

[0484] Exemplary bispecific antibodies may bind to two differentepitopes on a given PRO polypeptide herein. Alternatively, an anti-PROpolypeptide arm may be combined with an arm which binds to a triggeringmolecule on a leukocyte such as a T-cell receptor molecule (e.g. CD2,CD3, CD28, or B7), or Fc receptors for IgG (FcγR), such as FcγRI (CD64),FcγRII (CD32) and FcγRIII (CD16) so as to focus cellular defensemechanisms to the cell expressing the particular PRO polypeptide.Bispecific antibodies may also be used to localize cytotoxic agents tocells which express a particular PRO polypeptide. These antibodiespossess a PRO-binding arm and an arm which binds a cytotoxic agent or aradionuclide chelator, such as EOTUBE, DPTA, DOTA, or TETA. Anotherbispecific antibody of interest binds the PRO polypeptide and furtherbinds tissue factor (TF).

[0485] 5. Heteroconjugate Antibodies

[0486] Heteroconjugate antibodies are also within the scope of thepresent invention. Heteroconjugate antibodies are composed of twocovalently joined antibodies. Such antibodies have, for example, beenproposed to target immune system cells to unwanted cells [U.S. Pat. No.4,676,980], and for treatment of HIV infection [WO 91/00360; WO92/200373; EP 03089]. It is contemplated that the antibodies may beprepared in vitro using known methods in synthetic protein chemistry,including those involving crosslinking agents. For example, immunotoxinsmay be constructed using a disulfide exchange reaction or by forming athioether bond. Examples of suitable reagents for this purpose includeiminothiolate and methyl-4-mercaptobutyrimidate and those disclosed, forexample, in U.S. Pat. No. 4,676,980.

[0487] 6. Effector Function Engineering

[0488] It may be desirable to modify the antibody of the invention withrespect to effector function, so as to enhance, e.g., the effectivenessof the antibody in treating cancer. For example, cysteine residue(s) maybe introduced into the Fc region, thereby allowing interchain disulfidebond formation in this region. The homodimeric antibody thus generatedmay have improved internalization capability and/or increasedcomplement-mediated cell killing and antibody-dependent cellularcytotoxicity (ADCC). See Caron et al., J. Exp Med., 176: 1191-1195(1992) and Shopes, J. Immunol., 148: 2918-2922 (1992). Homodimericantibodies with enhanced anti-tumor activity may also be prepared usingheterobifunctional cross-linkers as described in Wolff et al. CancerResearch, 53: 2560-2565 (1993). Alternatively, an antibody can beengineered that has dual Fe regions and may thereby have enhancedcomplement lysis and ADCC capabilities. See Stevenson et al.,Anti-Cancer Drug: Design. 3: 219-230 (1989).

[0489] 7. Immunoconjugates

[0490] The invention also pertains to immunoconjugates comprising anantibody conjugated to a cytotoxic agent such as a chemotherapeuticagent, toxin (e.g., an enzymatically active toxin of bacterial, fungal,plant, or animal origin, or fragments thereof), or a radioactive isotope(i.e., a radioconjugate).

[0491] Chemotherapeutic agents useful in the generation of suchimmunoconjugates have been described above. Enzymatically active toxinsand fragments thereof that can be used include diphtheria A chain,nonbinding active fragments of diphtheria toxin, exotoxin A chain (fromPseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain,alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolacaamericana proteins (PAPI, PAPII, and PAP-S), momordica charantiainhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin,mitogellin, restrictocin, phenomycin, enomycin, and the tricothecenes. Avariety of radionuclides are available for the production ofradioconjugated antibodies. Examples include ²¹²Bi, ¹³¹I, ¹³¹In, ⁹⁰Y,and ¹⁸⁶Re. Conjugates of the antibody and cytotoxic agent are made usinga variety of bifunctional protein-coupling agents such asN-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), iminothiolane(IT), bifunctional derivatives of imidoesters (such as dimethyladipimidate HCL), active esters (such as disuccinimidyl suberate),aldehydes (such as glutareldehyde), bis-azido compounds (such as bis(p-azidobenzoyl) hexanediamine), bis-diazonium derivatives (such asbis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such astolyene 2,6-diisocyanate), and bis-active fluorine compounds (such as1,5-difluoro-2,4-dinitrobenzene). For example, a ricin immunotoxin canbe prepared as described in Vitetta et al., Science, 238: 1098 (1987).Carbon-14-labeled 1-isothiocyanatobenzyl-3-methyldiethylenetriaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent forconjugation of radionucleotide to the antibody. See W094/11026.

[0492] In another embodiment, the antibody may be conjugated to a“receptor” (such streptavidin) for utilization in tumor pretargetingwherein the antibody-receptor conjugate is administered to the patient,followed by removal of unbound conjugate from the circulation using aclearing agent and then administration of a “ligand” (e.g., avidin) thatis conjugated to a cytotoxic agent (e.g., a radionucleotide).

[0493] 8. Immunoliposomes

[0494] The antibodies disclosed herein may also be formulated asimmunoliposomes. Liposomes containing the antibody are prepared bymethods known in the art, such as described in Epstein et al., Proc.Natl. Acad. Sci. USA, 82: 3688 (1985); Hwang et al., Proc. Natl Acad.Sci. USA, 77: 4030 (1980); and U.S. Pat. Nos. 4,485,045 and 4,544,545.Liposomes with enhanced circulation time are disclosed in U.S. Pat. No.5,013,556.

[0495] Particularly useful liposomes can be generated by thereverse-phase evaporation method with a lipid composition comprisingphosphatidylcholine, cholesterol, and PEG-derivatizedphosphatidylethanolamine (PEG-PE). Liposomes are extruded throughfilters of defined pore size to yield liposomes with the desireddiameter. Fab′ fragments of the antibody of the present invention can beconjugated to the liposomes as described in Martin et al ., J. Biol.Chem., 257: 286-288 (1982) via a disulfide-interchange reaction. Achemotherapeutic agent (such as Doxorubicin) is optionally containedwithin the liposome. See Gabizon et al., J. National Cancer Inst.,81(19): 1484 (1989).

[0496] 9. Pharmaceutical Compositions of Antibodies

[0497] Antibodies specifically binding a PRO polypeptide identifiedherein, as well as other molecules identified by the screening assaysdisclosed hereinbefore, can be administered for the treatment of variousdisorders in the form of pharmaceutical compositions.

[0498] If the PRO polypeptide is intracellular and whole antibodies arcused as inhibitors, internalizing antibodies are preferred. However,lipofections or liposomes can also be used to deliver the antibody, oran antibody fragment, into cells. Where antibody fragments are used, thesmallest inhibitory fragment that specifically binds to the bindingdomain of the target protein is preferred. For example, based upon thevariable-region sequences of an antibody, peptide molecules can bedesigned that retain the ability to bind the target protein sequence.Such peptides can be synthesized chemically and/or produced byrecombinant DNA technology. See, e.g., Marasco et al., Proc. Natl. Acad.Sci. USA, 90: 7889-7893 (1993). The formulation herein may also containmore than one active compound as necessary for the particular indicationbeing treated, preferably those with complementary activities that donot adversely affect each other. Alternatively, or in addition, thecomposition may comprise an agent that enhances its function, such as,for example, a cytotoxic agent, cytokine, chemotherapeutic agent, orgrowth-inhibitory agent. Such molecules are suitably present incombination in amounts that are effective for the purpose intended.

[0499] The active ingredients may also be entrapped in microcapsulesprepared, for example, by coacervation techniques or by interfacialpolymerization, for example, hydroxymethylcellulose orgelatin-microcapsules and poly-(methylmethacylate) microcapsules,respectively, in colloidal drug delivery systems (for example,liposomes, albumin microspheres, microemulsions, nano-particles, andnanocapsules) or in macroemulsions. Such techniques are disclosed inRemington's Pharmaceutical Sciences, supra.

[0500] The formulations to be used for in vivo administration must besterile. This is readily accomplished by filtration through sterilefiltration membranes.

[0501] Sustained-release preparations may be prepared. Suitable examplesof sustained-release preparations include semipermeable matrices ofsolid hydrophobic polymers containing the antibody, which matrices arein the form of shaped articles, e.g., films, or microcapsules. Examplesof sustained-release matrices include polyesters, hydrogels (forexample, poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)),polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acidand γ ethyl-L-glutamate, non-degradable ethylene-vinyl acetate,degradable lactic acid-glycolic acid copolymers such as the LUPRONDEPOT™ (injectable microspheres composed of lactic acid-glycolic acidcopolymer and leuprolide acetate), and poly-D-(−)-3-hydroxybutyric acid.While polymers such as ethylene-vinyl acetate and lactic acid-glycolicacid enable release of molecules for over 100 days, certain hydrogelsrelease proteins for shorter time periods. When encapsulated antibodiesremain in the body for a long time, they may denature or aggregate as aresult of exposure to moisture at 37° C., resulting in a loss ofbiological activity and possible changes in immunogenicity. Rationalstrategies can be devised for stabilization depending on the mechanisminvolved. For example, if the aggregation mechanism is discovered to beintermolecular S-S bond formation through thio-disulfide interchange,stabilization may be achieved by modifying sulfliydryl residues,lyophilizing from acidic solutions, controlling moisture content, usingappropriate additives, and developing specific polymer matrixcompositions.

[0502] G. Uses for anti-PRO Antibodies

[0503] The anti-PRO antibodies of the invention have various utilities.For example, anti-PRO antibodies may be used in diagnostic assays forPRO, e.g., detecting its expression in specific cells, tissues, orserum. Various diagnostic assay techniques known in the art may be used,such as competitive binding assays, direct or indirect sandwich assaysand immunoprecipitation assays conducted in either heterogeneous orhomogeneous phases [Zola, Monoclonal Antibodies: A Manual of Techniques,CRC Press, Inc. (1987) pp. 147-158]. The antibodies used in thediagnostic assays can be labeled with a detectable moiety. Thedetectable moiety should be capable of producing, either directly orindirectly, a detectable signal. For example, the detectable moiety maybe a radioisotope, such as ³H, ¹⁴C, ³²p, ³⁵S, or ¹²⁵I, a fluorescent orchemiluminescent compound, such as fluorescein isothiocyanate,rhodamine, or luciferin, or an enzyme, such as alkaline phosphatase,beta-galactosidase or horseradish peroxidase. Any method known in theart for conjugating the antibody to the detectable moiety may beemployed, including those methods described by Hunter et al., Nature,144:945 (1962); David et al., Biochemistry, 13:1014 (1974); Pain et al.,J. Immunol. Meth 40:219 (1981); and Nygren, J. Histochem. and Cytochem.,30:407 (1982).

[0504] Anti-PRO antibodies also are useful for the affinity purificationof PRO from recombinant cell culture or natural sources. In thisprocess, the antibodies against PRO are immobilized on a suitablesupport, such a Sephadex resin or filter paper, using methods well knownin the art. The immobilized. antibody then is contacted with a samplecontaining the PRO to be purified, and thereafter the support is washedwith a suitable solvent that will remove substantially all the materialin the sample except the PRO, which is bound to the immobilizedantibody. Finally, the support is washed with another suitable solventthat will release the PRO from the antibody.

[0505] The following examples are offered for illustrative purposesonly, and are not intended to limit the scope of the present inventionin any way.

[0506] All patent and literature references cited in the presentspecification are hereby incorporated by reference in their entirety.

EXAMPLES

[0507] Commercially available reagents referred to in the examples wereused according to manufacturer's instructions unless otherwiseindicated. The source of those cells identified in the followingexamples, and throughout the specification, by ATCC accession numbers isthe American Type Culture Collection, Manassas, Va.

Example 1 Extracellular Domain Homology Screening to Identify NovelPolypeptides and cDNA Encoding Therefor

[0508] The extracellular domain (ECD) sequences (including the secretionsignal sequence, if any) from about 950 known secreted proteins from theSwiss-Prot public database were used to search EST databases. The ESTdatabases included public databases (e.g., Dayhoff, GenBank), andproprietary databases (e.g. LIFESEQ™, Incyte Pharmaceuticals, Palo Alto,Calif.). The search was performed using the computer program BLAST orBLAST-2 (Altschul et al., Methods in Enzymology 266:460-480 (1996)) as acomparison of the ECD protein sequences to a 6 frame translation of theEST sequences. Those comparisons with a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into consensus DNA sequences with the program “phrap”(Phil Green, University of Washington, Seattle, Wash.).

[0509] Using this extracellular domain homology screen, consensus DNAsequences were assembled relative to the other identified EST sequencesusing phrap. In addition, the consensus DNA sequences obtained wereoften (but not always) extended using repeated cycles of BLAST orBLAST-2 and phrap to extend the consensus sequence as far as possibleusing the sources of EST sequences discussed above.

[0510] Based upon the consensus sequences obtained as described above,oligonucleotides were then synthesized and used to identify by PCR acDNA library that contained the sequence of interest and for use asprobes to isolate a clone of the full-length coding sequence for a PROpolypeptide. Forward and reverse PCR primers generally range from 20 to30 nucleotides and are often designed to give a PCR product of about100-1000 bp in length. The probe sequences are typically 40-55 bp inlength. In some cases, additional oligonucleotides are synthesized whenthe consensus sequence is greater than about 1-1.5 kbp. In order toscreen several libraries for a full-length clone, DNA from the librarieswas screened by PCR amplification, as per Ausubel et al., CurrentProtocols in Molecular Biology, with the PCR primer pair. A positivelibrary was then used to isolate clones encoding the gene of interestusing the probe oligonucleotide and one of the primer pairs.

[0511] The cDNA libraries used to isolate the cDNA clones wereconstructed by standard methods using commercially available reagentssuch as those from Invitrogen, San Diego, Calif. The cDNA was primedwith oligo dT containing a NotI site, linked with blunt to SalIhemikinased adaptors, cleaved with NotI, sized appropriately by gelelectrophoresis, and cloned in a defined orientation into a suitablecloning vector (such as pRKB or pRKD; pRK5B is a precursor of pRK5D thatdoes not contain the SfiI site; see, Holmes et al., Science,253:1278-1280 (1991)) in the unique XhoI and NotI sites.

Example 2 Isolation of cDNA Clones by Amylase Screening

[0512] 1. Preparation of oligo dT primed cDNA library

[0513] mRNA was isolated from a human tissue of interest using reagentsand protocols from Invitrogen, San Diego, Calif. (Fast Track 2). ThisRNA was used to generate an oligo dT primed cDNA library in the vectorpRK5D using reagents and protocols from Life Technologies, Gaithersburg,Md. (Super Script Plasmid System). In this procedure, the doublestranded cDNA was sized to greater than 1000 bp and the SalI/NotIlinkered cDNA was cloned into XhoI/NotI cleaved vector. pRK5D is acloning vector that has an sp6 transcription initiation site followed byan SfiI restriction enzyme site preceding the XhoI/NotI cDNA cloningsites.

[0514] 2. Preparation of random primed cDNA library

[0515] A secondary cDNA library was generated in order to preferentiallyrepresent the 5′ ends of the primary cDNA clones. Sp6 RNA was generatedfrom the primary library (described above), and this RNA was used togenerate a random primed cDNA library in the vector pSST-AMY.0 usingreagents and protocols from Life Technologies (Super Script PlasmidSystem, referenced above). In this procedure the double stranded cDNAwas sized to 500-1000 bp, linkered with blunt to NotI adaptors, cleavedwith SfiI, and cloned into SfiI/NotI cleaved vector. pSST-AMY.0 is acloning vector that has a yeast alcohol dehydrogenase promoter precedingthe cDNA cloning sites and the mouse amylase sequence (the maturesequence without the secretion signal) followed by the yeast alcoholdehydrogenase terminator, after the cloning sites. Thus, cDNAs clonedinto this vector that are fused in frame with amylase sequence will leadto the secretion of amylase from appropriately transfected yeastcolonies.

[0516] 3. Transformation and Detection

[0517] DNA from the library described in paragraph 2 above was chilledon ice to which was added electrocompetent DH10B bacteria (LifeTechnologies, 20 ml). The bacteria and vector mixture was thenelectroporated as recommended by the manufacturer. Subsequently, SOCmedia (Life Technologies, 1 ml) was added and the mixture was incubatedat 37° C. for 30 minutes. The transformants were then plated onto 20standard 150 mn LB plates containing ampicillin and incubated for 16hours (37° C.). Positive colonies were scraped off the plates and theDNA was isolated from the bacterial pellet using standard protocols,e.g. CsCl-gradient. The purified DNA was then carried on to the yeastprotocols below.

[0518] The yeast methods were divided into three categories: (1)Transformation of yeast with the plasmid/cDNA combined vector; (2)Detection and isolation of yeast clones secreting amylase; and (3) PCRamplification of the insert directly from the yeast colony andpurification of the DNA for sequencing and further analysis.

[0519] The yeast strain used was HD56-5A (ATCC-90785). This strain hasthe following genotype: MAT alpha, ura3-52, leu2-3, leu2-112, his3-11,his3-15, MAL⁺, SUC⁺, GAL⁺. Preferably, yeast mutants can be employedthat have deficient post-translational pathways. Such mutants may havetranslocation deficient alleles in sec71, sec72, sec62, with truncatedsec71 being most preferred. Alternatively, antagonists (includingantisense nucleotides and/or ligands) which interfere with the normaloperation of these genes, other proteins implicated in this posttranslation pathway (e.g., SEC61p, SEC72p, SEC62p, SEC63p, TDJ1p orSSA1p-4p) or the complex formation of these proteins may also bepreferably employed in combination with the amylase-expressing yeast.

[0520] Transformation was performed based on the protocol outlined byGietz et al., Nucl. Acid. Res., 20:1425 (1992). Transformed cells werethen inoculated from agar into YEPD complex media broth (100 ml) andgrown overnight at 30° C. The YEPD broth was prepared as described inKaiser et al., Methods in Yeast Genetics, Cold Spring Harbor Press, ColdSpring Harbor, N.Y., p. 207 (1994). The overnight culture was thendiluted to about 2×10⁶ cells/ml (approx. OD₆₀₀=0.1) into fresh YEPDbroth (500 ml) and regrown to 1×10⁷ cells/ml (approx. OD₆₀₀=0.4-0.5).

[0521] The cells were then harvested and prepared for transformation bytransfer into GS3 rotor bottles in a Sorval GS3 rotor at 5,000 rpm for 5minutes, the supernatant discarded, and then resuspended into sterilewater, and centrifuged again in 50 ml falcon tubes at 3,500 rpm in aBeckman GS-6KR centrifuge. The supernatant was discarded and the cellswere subsequently washed with LiAc/TE (10 ml, 10 mM Tris-HCl, 1 mM EDTApH 7.5, 100 mM Li₂OOCCH₃), and resuspended into LiAc/TE (2.5 ml).

[0522] Transformation took place by mixing the prepared cells (100 μl)with freshly denatured single stranded salmon testes DNA (LofstrandLabs, Gaithersburg, Md.) and transforming DNA (1 μg, vol. <10 μl) inmicrofuge tubes. The mixture was mixed briefly by vortexing, then 40%PEG/TE (600 μl, 40% polyethylene glycol-4000, 10 mM Tris-HCl, 1 mM EDTA,100 mM Li₂OOCCH₃, pH 7.5) was added. This mixture was gently mixed andincubated at 30° C. while agitating for 30 minutes. The cells were thenheat shocked at 42° C. for 15 minutes, and the reaction vesselcentrifuged in a microfuge at 12,000 rpm for 5-10 seconds, decanted andresuspended into TE (500 μl, 10 mM Tris-HCl, 1 mM EDTA pH 7.5) followedby recentrifugation. The cells were then diluted into TE (1 ml) andaliquots (200 μl ) were spread onto the selective media previouslyprepared in 150 mm growth plates (VWR).

[0523] Alternatively, instead of multiple small reactions, thetransformation was performed using a single, large scale reaction,wherein reagent amounts were scaled up accordingly.

[0524] The selective media used was a synthetic complete dextrose agarlacking uracil (SCD-Ura) prepared as described in Kaiser et al., Methodsin Yeast Genetics, Cold Spring Harbor Press, Cold Spring Harbor, N.Y.,p. 208-210 (1994). Transformants were grown at 30° C. for 2-3 days.

[0525] The detection of colonies secreting amylase was performed byincluding red starch in the selective growth media. Starch was coupledto the red dye (Reactive Red-120, Sigma) as per the procedure describedby Biely et al., Anal. Biochem., 172:176-179 (1988). The coupled starchwas incorporated into the SCD-Ura agar plates at a final concentrationof 0.15% (w/v), and was buffered with potassium phosphate to a pH of 7.0(50-100 mM final concentration).

[0526] The positive colonies were picked and streaked across freshselective media (onto 150 mm plates) in order to obtain well isolatedand identifiable single colonies. Well isolated single colonies positivefor amylase secretion were detected by direct incorporation of redstarch into buffered SCD-Ura agar. Positive colonies were determined bytheir ability to break down starch resulting in a clear halo around thepositive colony visualized directly.

[0527] 4. Isolation of DNA by PCR Amplification

[0528] When a positive colony was isolated, a portion of it was pickedby a toothpick and diluted into sterile water (30 μl) in a 96 wellplate. At this time, the positive colonies were either frozen and storedfor subsequent analysis or immediately amplified. An aliquot of cells (5μl) was used as a template for the PCR reaction in a 25 μl volumecontaining: 0.5 μl Klentaq (Clontech, Palo Alto, Calif.); 4.0 μl 10 mMdNTP's (Perkin Elmer-Cetus); 2.5 μl Kentaq buffer (Clontech); 0.25 μlforward oligo 1; 0.25 μl reverse oligo 2; 12.5 μl distilled water. Thesequence of the forward oligonucleotide 1 was:

[0529] 5′-TGTAAAACGACGGCCAGTTAAATAGACCTGCAATTATTAATCT-3′ (SEQ ID NO: 1)

[0530] The sequence of reverse oligonucleotide 2 was:

[0531] 5′-CAGGAAACAGCTATGACCACCTGCACACCTGCAAATCCATT-3′ (SEQ ID NO: 2)

[0532] PCR was then performed as follows: a. Denature 92° C., 5 minutesb.  3 cycles of: Denature 92° C., 30 seconds Anneal 59° C., 30 secondsExtend 72° C., 60 seconds c.  3 cycles of: Denature 92° C., 30 secondsAnneal 57° C., 30 seconds Extend 72° C., 60 seconds d. 25 cycles of:Denature 92° C., 30 seconds Anneal 55° C., 30 seconds Extend 72° C., 60seconds e. Hold  4° C.

[0533] The underlined regions of the oligonucleotides annealed to theADH promoter region and the amylase region, respectively, and amplifieda 307 bp region from vector pSST-AMY.0 when no insert was present.Typically, the first 18 nucleotides of the 5′ end of theseoligonucleotides contained annealing sites for the sequencing primers.Thus, the total product of the PCR reaction from an empty vector was 343bp. However, signal sequence-fused cDNA resulted in considerably longernucleotide sequences.

[0534] Following the PCR, an aliquot of the reaction (5 μl) was examinedby agarose gel electrophoresis in a 1% agarose gel using aTris-Borate-EDTA (TBE) buffering system as described by Sambrook et al.,supra. clonesresulting in a single strong PCR product larger than 400 bpwere further analyzed by DNA sequencing after purification with a 96Qiaquick PCR clean-up column (Qiagen Inc., Chatsworth, Calif.).

Example 3 Isolation of cDNA Clones Using Signal Algorithm Analysis

[0535] Various polypeptide-encoding nucleic acid sequences wereidentified by applying a proprietary signal sequence finding algorithmdeveloped by Genentech, Inc. (South San Francisco, Calif.) upon ESTs aswell as clustered and assembled EST fragments from public (e.g.,GenBank) and/or private (LIFESEQ®, Incyte Pharmaceuticals, Inc., PaloAlto, Calif.) databases. The signal sequence algorithm computes asecretion signal score based on the character of the DNA nucleotidessurrounding the first and optionally the second methionine codon(s)(ATG) at the 5′-end of the sequence or sequence fragment underconsideration. The nucleotides following the first ATG must code for atleast 35 unambiguous amino acids without any stop codons. If the firstATG has the required amino acids, the second is not examined. If neithermeets the requirement, the candidate sequence is not scored. In order todetermine whether the EST sequence contains an authentic signalsequence, the DNA and corresponding amino acid sequences surrounding theATG codon are scored using a set of seven sensors (evaluationparameters) known to be associated with secretion signals. Use of thisalgorithm resulted in the identification of numerouspolypeptide-encoding nucleic acid sequences.

Example 4 Isolation of cDNA Clones Encoding Human PRO196

[0536] PRO196 was identified by screening the GenBank database using thecomputer program BLAST (Altshul et al., Methods in Enzymology266:460-480 (1996). The PRO196 sequence shows homology with knownexpressed sequence tag (EST) sequences T35448, T11442, and W77823. Noneof the known EST sequences have been identified as full lengthsequences, or described as ligands associated with the TIE receptors.

[0537] Following its identification, NL1 was cloned from a human fetallung library prepared from mRNA purchased from Clontech, Inc. (PaloAlto, Calif., USA), catalog # 6528-1, following the manufacturer'sinstructions. The library was screened by hybridization with syntheticoligonucleotide probes:

[0538] (a) 5′-GCTGACGAACCAAGGCAACTACAAACTCCTGGT-3′ (SEQ ID NO: 5);

[0539] (b) 5′-TGCGGCCGGACCAGTCCTCCATGGTCACCAGGAGTTTGTAG-3′ (SEQ ID NO:6);

[0540] (c) 5′-GGTGGTGAACTGCTTGCCGTTGTGCCATGTAAA-3′ (SEQ ID NO: 7).

[0541] based on the ESTs found in the GenBank database. cDNA sequenceswere sequenced in their entireties.

[0542] The nucleotide and amino acid sequences of PRO196 are shown inFIG. 1 (SEQ ID NO: 3) and FIG. 2 (SEQ ID NO: 4), respectively. PRO196shows significant sequence identity with both the TIE1 and the TIE2ligand.

[0543] A clone of PRO196 was deposited with the American Type CultureCollection, 10801 University Blvd., Manassas, Va. 20110-2209, USA (ATCC)on Sep. 18, 1997 under the terms of the Budapest Treaty, and has beenassigned the deposit number 209280.

Example 5 Isolation of cDNA Clones Encoding Human PRO444

[0544] A cDNA sequence isolated in the amylase screen described inExample 2 above was designated DNA13121. Oligonucleotide probes weregenerated to this sequence and used to screen a human fetal lung library(LIB25) prepared as described in paragraph 1 of Example 2 above. Thecloning vector was pRK5B (pRK5B is a precursor of pRK5D that does notcontain the SfiI site; see, Holmes et al., Science, 253:1278-1280(1991)), and the cDNA size cut was less than 2800 bp.

[0545] A full length clone was identified that contained a single openreading frame with an apparent translational initiation site atnucleotide positions 608-610 and ending at the stop codon found atnucleotide positions 959-961 (FIG. 3, SEQ ID NO: 8). The predictedpolypeptide precursor is 117 amino acids long, has a calculatedmolecular weight of approximately 12,692 daltons and an estimated pI ofapproximately 7.50. Analysis of the full-length PRO444 sequence shown inFIG. 4 (SEQ ID NO: 9) evidences the presence of a signal peptide atamino acid 1 to about amino acid 16. An analysis of the Dayhoff database(version 35.45 SwissProt 35) evidenced homology between the PRO444 aminoacid sequence and the following Dayhoff sequences: CEF44D12_(—)8,P_R88452, YNE1_CAEEL, A47312, AF009957_(—)1, and A06133_(—)1.

[0546] Clone DNA26846-1397 was deposited with the ATCC on Oct. 27, 1998and is assigned ATCC deposit no. 203406.

Example 6 Isolation of cDNA Clones Encoding Human PRO183, PRO185,PRO9940, PRO2630 and PRO6309

[0547] DNA molecules encoding the PRO183, PRO185, PRO9940, PRO2630 andPRO6309 polypeptides shown in the accompanying figures were obtainedthrough GenBank.

Example 7 Isolation of cDNA Clones Encoding Human PRO210 and PRO217

[0548] A consensus DNA sequence was assembled using phrap as describedin Example 1 above. In some cases, the consensus DNA sequence asextended using repeated cycles of blast and phrap to extend theconsensus sequence as far as possible using the sources of EST sequenceslisted above. Based on this consensus sequence, oligonucleotides weresynthesized: 1) to identify by PCR a cDNA library that contained thesequence of interest, and 2) for use as probes to isolate a clone of thefull-length coding sequence. The library used to isolate DNA32279-1131was fetal kidney.

[0549] cDNA clones were sequenced in their entirety. The entirenucleotide sequence of DNA32279-1131 is shown in FIG. 9 (SEQ ID NO: 14)and amino acid sequence of PRO210 is shown in FIG. 10 (SEQ ID NO: 15).The entire nucleotide sequence of DNA33094-1131 is shown in FIG. 13 (SEQID NO: 21) and amino acid sequence of PRO217 is shown in FIG. 14 (SEQ IDNO: 22).

Example 8 Isolation of cDNA clones Encoding Human PRO215

[0550] A consensus DNA sequence was assembled relative to the otheridentified EST sequences as described in Example 1 above, wherein theconsensus sequence was designated herein as DNA28748. Based on theDNA28748 consensus sequence, oligonucleotides were synthesized toidentify by PCR a cDNA library that contained the sequence of interestand for use as probes to isolate a clone of the full-length codingsequence for PRO215.

[0551] A pair of PCR primers (forward and reverse) were synthesized:

[0552] forward PCR primer 5′-GTGGCTGGAAAATGAGATC-3′ (SEQ ID NO: 18)

[0553] reverse PCR primer 5′-CAATGTGTGAAGCGGTTGTG-3′ (SEQ ID NO: 19)

[0554] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA28748 sequence which had the followingnucleotide sequence:

[0555] hybridization probe

[0556] 5′-TAAGAGCCTGGACCTAGCAAATCTATCTCTGACTTTGCCTGGAGC-3 (SEQ ID NO:20).

[0557] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO215 gene usingthe probe oligonucleotide and one of the PCR primers.

[0558] RNA for construction of the cDNA libraries was isolated fromhuman fetal lung tissue. The cDNA libraries used to isolate the cDNAclones were constructed by standard methods using commercially availablereagents such as those from Invitrogen, San Diego, Calif. The cDNA wasprimed with oligo dT containing a NotI site, linked with blunt to SalIhemikinased adaptors, cleaved with NotI, sized appropriately by gelelectrophoresis, and cloned in a defined orientation into a suitablecloning vector (such as pRKB or pRKD; pRK5B is a precursor of pRK5D thatdoes not contain the SfiI site; see, Holmes et al., Science,253:1278-1280 (1991)) in the unique XhoI and NotI sites.

[0559] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO215 [herein designated as DNA32288-1132] and the derived protein sequence for PRO215.

[0560] The entire nucleotide sequence of DNA32288-1132 is shown in FIG.11 (SEQ ID NO: 16). Clone DNA32288-1132 contains a single open readingframe with an apparent translational initiation site at nucleotidepositions 308-310 and ending at the stop codon at nucleotide positions1591-1593 (FIG. 11, the initiation and stop codons are circled). Thepredicted polypeptide precursor is 428 amino acids long (FIG. 12). CloneDNA32288-1132 has been deposited with ATCC and is assigned ATCC depositno. 209261.

[0561] Analysis of the amino acid sequence of the full-length PRO215shows it has homology to member of the leucine rich repeat proteinsuperfamily, including the leucine rich repeat protein and the SLITprotein.

Example 9 Isolation of cDNA Clones Encoding Human PRO242

[0562] An expressed sequence tag (EST) DNA database (LIFESEQ™, IncytePharmaceuticals, Palo Alto, Calif.) was searched and an EST wasidentified which showed homology to a chemokine. Based on this sequence,oligonucleotides were synthesized to identify by PCR a cDNA library thatcontained the sequence of interest and for use as probes to isolate aclone of the full-length coding sequence for PRO242.

[0563] A pair of PCR primer (forward and reverse) were synthesized:

[0564] forward PCR primer 5′-GGATAGGAGGAGGAGTTTGGG-3′ (SEQ ID NO: 25)

[0565] reverse PCR primer 5′-GGATGGGTAAGACTTTCTTGCC-3′ (SEQ ID NO: 26)

[0566] Additionaly, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA28709 sequence which had the followingnucleotide sequence:

[0567] hybridization probe

[0568] 5′-ATGATGGGCCTCTCCTTGGCCTCTGCTGTGCTCCTGGCCTCCCTCCTGAG-3-(SEQ IDNO: 27)

[0569] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO242 gene usingthe probe oligonucleotide and one of the PCR primers.

[0570] RNA for construction of the cDNA libraries was isolated fromhuman fetal lung tissue. A cDNA clone was sequenced in entirety. Theentire nucleotide sequence of DNA33785-1143 is shown in FIG. 15 (SEQ IDNO: 23). Clone DNA33785-1143 contains a single open reading frame withan apparent translational initiation site at nucleotide positions333-335 and ending at the stop codon at nucleotide positions 615-617(FIG. 16; SEQ ID NO: 24). The predicted polypeptide precursor is 94amino acids long (FIG. 16).

[0571] Based on a BLAST and FastA sequence alignment analysis (using theALIGN computer program) of the full-length sequence, PRO242 shows aminoacid sequence identity to human macrophage inflammatory protein 1-alpha,rabbitt macrophage inflammatory protein 1-beta, human LD78 and rabbitimmune activation gene 2.

Example 10 Isolation of cDNA Clones Encoding Human PRO288

[0572] A synthetic probe based on the sequence encoding the DcR1 ECD[Sheridan et al., supral] and having the following sequence:

[0573] 5′-CATAAAAGTTCCTGACACCATGACCAGAGACACATGTGTCAGTGTAAAGA-3′ (SEQ IDNO: 30)

[0574] was used to screen a human fetal lung cDNA library. To preparethe cDNA library, mRNA was isolated from human fetal lung tissue usingreagents and protocols from Invitrogen, San Diego, Calif. (Fast Track2). This RNA was used to generate an oligo dT primed cDNA library in thevector pRK5D using reagents and protocols from Life Technologies,Gaithersburg, Md. (Super Script Plasmid System). In this procedure, thedouble stranded cDNA was sized to greater than 1000 bp and the SalI/NotIlinkered cDNA was cloned into XhoI/NotI cleaved vector. pRK5D is acloning vector that has an sp6 transcription initiation site followed byan SfiI restriction enzyme site preceding the XhoI/NotI cDNA cloningsites.

[0575] A full length clone was identified (DNA35663-1129) that containeda single open reading frame with an apparent translational initiationsite at nucleotide positions 157-159 and ending at the stop codon foundat nucleotide positions 1315-1317 (FIG. 17; SEQ ID NO: 28). The clone isreferred to as pRK5-35663 and is deposited as ATCC No. 209201.

[0576] The predicted polypeptide precursor is 386 amino acids long andhas a calculated molecular weight of approximately 41.8 kDa. Sequenceanalysis indicated a N-terminal signal peptide (amino acids 1-55),followed by an ECD (amino acids 56-212), transmembrane domain (aminoacids 213-232) and intracellular region (amino acids 233-386). (FIG.18). The signal peptide cleavage site was confirmed by N-terminalprotein sequencing of a PRO288 ECD immunoadhesin (not shown). Thisstructure suggests that PRO288 is a type I transmembrane protein. PRO288contains 3 potential N-linked glycosylation sites, at amino acidpositions 127, 171 and 182. (FIG. 18)

[0577] TNF receptor family proteins are typically characterized by thepresence of multiple (usually four) cysteine-rich domains in theirextracellular regions—each cysteine-rich domain being approximately 45amino acids long and containing approximately 6, regularly spaced,cysteine residues. Based on the crystal structure of the type 1 TNFreceptor, the cysteines in each domain typically form three disulfidebonds in which usually cysteines 1 and 2, 3 and 5, and 4 and 6 arepaired together. Like DR4, DR5, and DcR1, PRO288 contains twoextracellular cysteine-rich pseudorepeats, whereas other identifiedmammalian TNFR family members contain three or more such domains [Smithet al., Cell, 76:959 (1994)].

[0578] Based on an alignment analysis of the PRO288 sequence shown inFIG. 18 (SEQ ID NO: 29), PRO288 shows more sequence identity to the ECDof DR4, DR5, or DcR1 than to other apoptosis-linked receptors, such asTNFR1, Fas/Apo-1 or DR3. The predicted intracellular sequence of PRO288also shows more homology to the corresponding region of DR4 or DR5 ascompared to TNFR1, Fas or DR3. The intracellular region of PRO288 isabout 50 residues shorter than the intracellular regions identified forDR4 or DR5. It is presently believed that PRO288 may contain antruncated death domain (amino acids 340-364), which corresponds to thecarboxy-terminal portion of the death domain sequences of DR4 and DR5.Five out of six amino acids that are essential for signaling by TNFR1[Tartaglia et al., supra] and that are conserved or semi-conserved inDR4 and DR5, are absent in PRO288.

Example 11 Isolation of cDNA Clones Encoding Human PRO365

[0579] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequence is herein designated DNA35613. Based on the DNA35613 consensussequence, oligonucleotides were synthesized: 1) to identify by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence for PRO365.

[0580] Forward and reverse PCR primers were synthesized:

[0581] forward PCR primer 5′-GGCTGGCCTGCAGAGATC-3′ (SEQ ID NO: 33)

[0582] forward PCR primer 5′-AATGTGACCACTGGACTCCC-3′ (SEQ ID NO: 34)

[0583] forward PCR primer 5′-AGGCTTGGAACTCCCTTC-3′ (SEQ ID NO: 35)

[0584] reverse PCR primer 5′-AAGATTCTTGAGCGATTCCAGCTG-3′ (SEQ ID NO: 36)

[0585] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA35613 sequence which had the followingnucleotide sequence

[0586] hybridization probe

[0587] 5 ′-AATCCCTGCTCTTATGGTGACCTCATGACGACGGAAGCAAAGCACTG-3′ (SEQ IDNO: 37)

[0588] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO365gene using the probe oligonucleotide and one of the PCR primers.RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue.

[0589] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO365 [herein designated as DNA46777-1253](SEQ ID NO: 31) and the derived protein sequence for PRO365.

[0590] The entire nucleotide sequence of DNA46777-1253 is shown in FIG.19 (SEQ ID NO: 31). Clone DNA46777-1253 contains a single open readingframe with an apparent translational initiation site at nucleotidepositions 15-17 and ending at the stop codon at nucleotide positions720-722 (FIG. 19). The predicted polypeptide precursor is 235 aminoacids long (FIG. 20). Important regions of the polypeptide sequenceencoded by clone DNA46777-1253 have been identified and include thefollowing: a signal peptide corresponding to amino acids 1-20 andmultiple potential N-glycosylation sites. Clone DNA46777-1253 has beendeposited with ATCC and is assigned ATCC deposit no. 209619.

[0591] Analysis of the amino acid sequence of the full-length PRO365polypeptide suggests that portions of it possess significant homology tothe human 2-19 protein, thereby indicating that PRO365 may be a novelhuman 2-19 protein homolog.

Example 12 Isolation of cDNA Clones Encoding Human PRO1361

[0592] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from the Incytedatabase, designated Incyte cluster sequence 10685. This EST clustersequence was then compared to a variety of expressed sequence tag (EST)databases which included public EST databases (e.g., GenBank) and aproprietary EST DNA database (Lifeseq®, Incyte Pharmaceuticals, PaloAlto, Calif.) to identify existing homologies. The homology search wasperformed using the computer program BLAST or BLAST2 (Altshul et al.,Methods in Enzymolopy 266:460-480 (1996)). Those comparisons resultingin a BLAST score of 70 (or in some cases 90) or greater that did notencode known proteins were clustered and assembled into a consensus DNAsequence with the program “phrap” (Phil Green, University of Washington,Seattle, Wash.). The consensus sequence obtained therefrom is hereindesignated DNA58839.

[0593] In light of an observed sequence homology between the DNA58839sequence and an EST sequence contained within the Incyte EST clone no.2967927, the Incyte EST clone no. 2967927 was purchased and the cDNAinsert was obtained and sequenced. The sequence of this cDNA insert isshown in FIG. 21 and is herein designated as DNA60783-1611.

[0594] Clone DNA60783-1611 contains a single open reading frame with anapparent translational initiation site at nucleotide positions 142-144and ending at the stop codon at nucleotide positions 1132-1134 (FIG.21). The predicted polypeptide precursor is 330 amino acids long (FIG.22). The full-length PRO1361 protein shown in FIG. 22 has an estimatedmolecular weight of about 36,840 daltons and a pI of about 4.84.Analysis of the full-length PRO1361 sequence shown in FIG. 22 (SEQ IDNO: 39) evidences the presence of the following: a signal peptide fromabout amino acid 1 to about amino acid 23, a transmembrane domain fromabout amino acid 266 to about amino acid 284, a leucine zipper patternsequence from about amino acid 155 to about amino acid 176 and potentialN-glycosylation sites from about amino acid 46 to about amino acid 49,from about amino acid 64 to about amino acid 67, from about amino acid166 to about amino acid 169 and from about amino acid 191 to about aminoacid 194. Clone DNA60783-1611 has been deposited with ATCC on Aug. 18,1998 and is assigned ATCC deposit no. 203130.

[0595] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 22 (SEQ ID NO: 39), evidenced significanthomology between the PRO1361 amino acid sequence and the followingDayhoff sequences: I50620, G64876, PMCMSG102B_(—)2MSG104, HUMIGLVXY_(—)1and PH1370.

Example 13 Isolation of cDNA Clones Encoding Human PRO1308

[0596] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. The consensussequence was extended then using repeated cycles of BLAST and phrap toextend the consensus sequence as far as possible using the sources ofEST sequences discussed above. The extended consensus sequence isdesignated herein as “DNA35726”. Based on the DNA35726 consensussequence, oligonucleotides were synthesized: 1) to identify by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence forPRO1308.

[0597] The following PCR primers (forward and reverse) were synthesized:

[0598] forward PCR primers 5′-TCCTGTGAGCACGTGGTGTG-3′ (SEQ ID NO: 42);

[0599] 5′-GGGTGGGATAGACCTGCG-3′ (SEQ ID NO: 43);

[0600] 5′-AAGGCCAAGAAGGCTGCC-3′ (SEQ ID NO: 44); and

[0601] 5′-CCAGGCCTGCAGACCCAG-3′ (SEQ ID NO: 45).

[0602] reverse PCR primers 5′-CTTCCTCCAGTCCTTCCAGGATATC-3′ (SEQ ID NO:46);

[0603] 5′-AAGCTGGATATCCTCCGTGTTGTC-3′ (SEQ ID NO: 47);

[0604] 5′-CCTGAAGAGGATGCACTGCTTTTCTCA-3′ (SEQ ID NO: 48); and

[0605] 5′-GGGGATAAACCTATTAATTATTGCTAC-3′ (SEQ ID NO: 49).

[0606] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA35726 sequence which had the followingnucleotide sequence: hybridization probe:5′-AACGTCACCTACATCTCCTCGTGCCACATGCGCCAGGCCACCTG-3′ (SEQ ID NO:50).

[0607] In order to screen several libraries for a source of full-lengthclone, DNA from the libraries was screened by PCR amplification with thePCR primer pair identified above. A positive library was then used toisolate clones encoding the PRO1308 gene using the probe oligonucleotideand one of the PCR primers. RNA for construction of the cDNA librarieswas isolated from a human SK-Lu-1 adenocarcinoma cell line.

[0608] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO1308 (designated herein as DNA62306-1570[FIG. 23, SEQ ID NO: 40]; and the derived protein sequence for PRO1308.

[0609] The entire coding sequence of PRO1308 is shown in FIG. 23 (SEQ IDNO: 40). Clone DNA62306-1570 contains a single open reading frame withan apparent translational initiation site at nucleotide positions 17-19and an apparent stop codon at nucleotide positions 806-808. Thepredicted polypeptide precursor is 263 amino acids long. The full-lengthPRO1308 protein shown in FIG. 24 has an estimated molecular weight ofabout 27,663 daltons and a pI of about 6.77. Additional features includea signal peptide at about amino acids 1-20, potential N-glycosylationsites at about amino acids 73-76 and 215-218, and regions of homologywith osteonectin domains at about amino acids 97-129 and 169-201.

[0610] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 24 (SEQ ID NO: 41), revealed significant homologybetween the PRO1308 amino acid sequence and Dayhoff sequence S55369.Homology was also revealed between the PRO1308 amino acid sequence andthe following Dayhoff sequences: FSA_HUMAN, P_R20063, CELT13C2_(—)1,AGRI_RAT, p_W09406, G01639, SC1_(—RAT, S)60062, S51362, and IOV7_CHICK.

[0611] Clone DNA62306-1570 has been deposit with ATCC and is assignedATCC deposit no. 203254.

Example 14 Isolation of cDNA Clones Encoding Human PRO1183

[0612] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from the Incytedatabase. This EST cluster sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., GenBank) and a proprietary EST DNA database (LIFESEQ®,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify existinghomologies. The homology search was performed using the computer programBLAST or BLAST2 (Altshul et al., Methods in Enzymology 266:460-480(1996)). Those comparisons resulting in a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into a consensus DNA sequence with the program “phrap”(Phil Green, University of Washington, Seattle, Wash.). The consensussequence obtained therefrom is herein designated DNA56037.

[0613] In light of an observed sequence homology between the DNA56037sequence and an EST sequence contained within the Incyte EST 1645856(from a library constructed from prostate tumor tissue), the clone whichincludes EST 1645856 was purchased and the cDNA insert was obtained andsequenced. The sequence of this cDNA insert is shown in FIG. 25 and isherein designated as DNA62880-1513.

[0614] The full length clone shown in FIG. 25 contained a single openreading frame with an apparent translational initiation site atnucleotide positions 20-22 and ending at the stop codon found atnucleotide positions 1535-1537 (FIG. 25; SEQ ID NO: 51). The predictedpolypeptide precursor (FIG. 26, SEQ ID NO: 52) is 505 amino acids long.The signal peptide is approximately at amino acids 1-23 of SEQ ID NO:52. PRO1183 has a calculated molecular weight of approximately 56,640daltons and an estimated pI of approximately 6.1. Clone DNA62880-1513was deposited with the ATCC on Aug. 4, 1998 and is assigned ATCC depositno. 203097.

[0615] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 26 (SEQ ID NO: 52), revealed sequence identitybetween the PRO1183 amino acid sequence and the following Dayhoffsequences: MTV010_(—)1, P_W41604, S54021, AOFB_HUMAN, NPAJ4683_(—)1,S74689, GEN13608, ACHC_ACHFU, AB011173_(—)1 and PUO_MICRU. It isbelieved that administration of PRO1183 or regulators thereof may treatcertain oxidase disorders such as variegate porphyria.

Example 15 Isolation of cDNA Clones Encoding Human PRO1272

[0616] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from the Incytedatabase. This EST cluster sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., GenBank) and a proprietary EST DNA database (LIFESEQ®,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify existinghomologies. The homology search was performed using the computer programBLAST or BLAST2 (Altshul et al., Methods in Enzymology 266:460-480(1996)). Those comparisons resulting in a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into a consensus DNA sequence with the program “phrap”(Phil Green, University of Washington, Seattle, Wash.). The consensussequence obtained therefrom is herein designated DNA58753.

[0617] In light of an observed sequence homology between the DNA58753sequence and an EST sequence contained witin the EST clone 3049165, theIncyte clone (from a lung library) including EST 3049165 was purchasedand the cDNA insert was obtained and sequenced. The sequence of thiscDNA insert is shown in FIG. 27 and is herein designated asDNA64896-1539.

[0618] The full length clone shown in FIG. 27 contained a single openreading frame with an apparent translational initiation site atnucleotide positions 58-60 and ending at the stop codon found atnucleotide positions 556-558 (FIG. 27; SEQ ID NO: 53). The predictedpolypeptide precursor (FIG. 28, SEQ ID NO: 54) is 166 amino acids long.The signal peptide is at about amino acids 1-23 of SEQ ID NO: 54.PRO1272 has a calculated molecular weight of approximately 19,171daltons and an estimated pI of approximately 8.26. Clone DNA64896-1539was deposited with the ATCC on Sep. 9, 1998 and is assigned ATCC depositno. 203238.

[0619] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 28 (SEQ ID NO: 54), revealed sequence identitybetween the PRO1272 amino acid sequence and the following Dayhoffsequences (information from database incorporated herein): AF025474_,D69100, AE000757_(—)10, H69466, CELC50E3_(—)12, XLRANBP1_(—)1,YD67_SCHPO, B69459, H36856, and FRU40755_(—)1.

Example 16 Isolation of cDNA clones Encoding Human PRO1419

[0620] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from the Incytedatabase. This EST cluster sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., GenBank) and a proprietary EST DNA database (LIFESEQ®,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify existinghomologies. One or more of the ESTs was derived from a diseased tonsiltissue library. The homology search was performed using the computerprogram BLAST or BLAST2 (Altshul et al., Methods in Enzymology266:460-480 (1996)). Those comparisons resulting in a BLAST score of 70(or in some cases 90) or greater that did not encode known proteins wereclustered and assembled into a consensus DNA sequence with the program“phrap” (Phil Green, University of Washington, Seattle, Wash.). Theconsensus sequence obtained therefrom is herein designated DNA59761.

[0621] In light of an observed sequence homology between the DNA59761sequence and an EST sequence contained within the Incyte EST 3815008,the clone including this EST was purchased and the cDNA insert wasobtained and sequenced. The sequence of this cDNA insert is shown inFIG. 29 and is herein designated as DNA71290-1630.

[0622] The full length clone shown in FIG. 29 contained a single openreading frame with an apparent translational initiation site atnucleotide positions 86-88 and ending at the stop codon found atnucleotide positions 341-343 (FIG. 29; SEQ ID NO: 55). The predictedpolypeptide precursor (FIG. 30, SEQ ID NO: 56) is 85 amino acids longwith the signal peptide at about amino acids 1-17 of SEQ ID NO: 56.PRO1419 has a calculated molecular weight of approximately 9,700 daltonsand an estimated pI of approximately 9.55. Clone DNA71290-1630 wasdeposited with the ATCC on Sep. 22, 1998 and is assigned ATCC depositno. 203275.

[0623] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 30 (SEQ ID NO: 56), revealed sequence identitybetween the PRO1419 amino acid sequence and the following Dayhoffsequences (data incorporated herein): S07975 (B3-hordein), C48232,HOR7_HORVU, GEN11764, S14970, AF020312_(—)1, STAJ3220_(—)1,CER07E3_(—)1, CEY37A1B_(—)4, and ATAC00423810.

Example 17 Isolation of cDNA Clones Encoding Human PRO4999

[0624] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequence is herein designated DNA86634. Based on the DNA86634 consensussequence, oligonucleotides were synthesized: 1) to identify by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence forPRO4999.

[0625] PCR primers (forward and reverse) were synthesized:

[0626] forward PCR primer 5′-CCACTTGCCATGAACATGCCAC-3′ (SEQ ID NO: 59)

[0627] reverse PCR primer 5′-CCTCTTGACAGACATAGCGAGCCAC-3′ (SEQ ID NO:60)

[0628] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA86634 sequence which had the followingnucleotide sequence

[0629] hybridization probe

[0630] 5′-CACTCTTGTCTGTGGGAACCACACATCTTGCCACAACTGTGGC-3′ (SEQ ID NO: 61)

[0631] RNA for construction of the cDNA libraries was isolated fromhuman testis tissue. DNA sequencing of the clones isolated as describedabove gave the full-length DNA sequence for a full-length PRO4999polypeptide (designated herein as DNA96031-2664 [FIG. 31, SEQ ID NO:57]) and the derived protein sequence for that PRO4999 polypeptide.

[0632] The full length clone identified above contained a single openreading frame with an apparent translational initiation site atnucleotide positions 42-44 and a stop signal at nucleotide positions2283-2285 (FIG. 31, SEQ ID NO: 57). The predicted polypeptide precursoris 747 amino acids long, has a calculated molecular weight ofapproximately 82,710 daltons and an estimated pI of approximately 6.36.Analysis of the full-length PRO4999 sequence shown in FIG. 32 (SEQ IDNO: 58) evidences the presence of a variety of important polypeptidedomains as shown in FIG. 32, wherein the locations given for thoseimportant polypeptide domains are approximate as described above. CloneDNA96031-2664 has been deposited with ATCC on Jun. 15, 1999 and isassigned ATCC deposit no. 237-PTA.

[0633] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using the ALIGN-2 sequence alignment analysis of the full-lengthsequence shown in FIG. 32 (SEQ ID NO: 58), evidenced sequence identitybetween the PRO4999 amino acid sequence and the following Dayhoffsequences: UROM_HUMAN; FBN1_HUMAN; GGU88872_(—)1; S52111; GEN12408;P_R79478; P_W48756; P_R53087; P_R14584; and S78549.

Example 18 Isolation of cDNA Clones Encoding Human PRO7170

[0634] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from the LIFESEQ®database, Incyte Pharmaceuticals, Palo Alto, designated herein asCLU57836. This EST cluster sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., Genbank) and a proprietary EST DNA database (LIFESEQ®,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify existinghomologies. The homology search was performed using the computer programBLAST or BLAST2 (Altshul et al., Methods in Enzymology 266:460-480(1996)). Those comparisons resulting in a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into a consensus DNA sequence with the program “phrap”(Phil Green, University of Washington, Seattle, Wash.). The consensussequence obtained therefrom is herein designated DNA58756.

[0635] In light of an observed sequence homology between the DNA58756sequence and an EST sequence encompassed within clone no. 2251462 fromthe LIFESEQ® database, Incyte Pharmaceuticals, Palo Alto, Calif., cloneno. 2251462 was purchased and the cDNA insert was obtained andsequenced. It was found herein that that cDNA insert encoded afull-length protein. The sequence of this cDNA insert is shown in FIG.33 and is herein designated as DNA108722-2743.

[0636] Clone DNA108722-2743 contains a single open reading frame with anapparent translational initiation site at nucleotide positions 60-62 andending at the stop codon at nucleotide positions 1506-1508 (FIG. 33).The predicted polypeptide precursor is 482 amino acids long (FIG. 34).The full-length PRO7170 protein shown in FIG. 34 has an estimatedmolecular weight of about 49,060 daltons and a pI of about 4.74.Analysis of the full-length PRO7170 sequence shown in FIG. 34 (SEQ IDNO: 63) evidences the presence of a variety of important polypeptidedomains as shown in FIG. 34, wherein the locations given for thoseimportant polypeptide domains are approximate as described above. CloneDNA108722-2743 has been deposited with ATCC on Aug. 17, 1999 and isassigned ATCC Deposit No. 552-PTA.

[0637] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using the ALIGN-2 sequence alignment analysis of the full-lengthsequence shown in FIG. 34 (SEQ ID NO: 63), evidenced sequence identitybetween the PRO7170 amino acid sequence and the following Dayhoffsequences: P_Y12291, I47141, D88733_(—)1, DMC56G7_(—)1, P_Y11606,HWP1_CANAL, HSMUC5BEX_(—)1, HSU78550_(—)1, HSU70136_(—)1, andSGS3_DROME.

Example 19 Isolation of cDNA Clones Encoding Human PRO248

[0638] A consensus DNA sequence was assembled relative to the otheridentified EST sequences as described in Example 1 above, wherein theconsensus sequence is designated herein as DNA33481. Based on theDNA33481 consensus sequence, oligonucleotides were synthesized toidentify by PCR a cDNA library that contained the sequence of interestand for use as probes to isolate a clone of the full-length codingsequence for PRO248. Specifically, the following primers were used:

[0639] Forward primer 1 (SEO ID NO: 66): 5′-GTCTGACAGCCACTCCAGAG-3′Hybridization probe (SEQ ID NO:67):5′-TCTCCAATTTCTGGGCTTAGATAAGGCGCCTTCACCCCAGAAGTTCC-3′

[0640] Reverse primer 1 (SEO ID NO: 68): 5′-GTCCCAGGTTATAGTAAGAATTGG-3′

[0641] Forward primer 2 (SEO ID NO: 69): 5′-GTGTTGCGGTAGTCCCATG-3′

[0642] Forward primer 3 (SEQ ID NO: 70): 5′-GCTGTCTCCCATTTCCATGC-3′

[0643] Reverse primer 2 (SEO ID NO: 71): 5′-CGACTACCATGTCTTCATAATGTC-3′

[0644] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO248 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue.

[0645] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO248 [herein designated as DNA35674-1142]and the derived protein sequence for PRO248.

[0646] The entire nucleotide sequence of DNA35674-1142 is shown in FIG.35 (SEQ ID NO: 64). Clone DNA35674-1142 contains a single open readingframe with an apparent translational initiation site at nucleotidepositions 66-68 and ending at the stop codon at nucleotide positions1217-1219 (FIG. 35; SEQ ID NO: 64). The predicted polypeptide precursoris 364 amino acids long (FIG. 36). Clone DNA35674-1142 has beendeposited on Oct. 28, 1997 with ATCC and is assigned ATCC deposit no.209416.

[0647] Analysis of the amino acid sequence of the full-length PRO248suggests that it has certain amino acid sequence identity with growthdifferentiation factor 3 from human and mouse.

Example 20 Isolation of cDNA Clones Encoding Human PRO353

[0648] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequences is herein designated DNA36363. The consensus DNA sequence wasextended using repeated cycles of BLAST and phrap to extend theconsensus sequence as far as possible using the sources of EST sequencesdiscussed above. Based on the DNA36363 consensus sequence,oligonucleotides were synthesized: 1) to identify by PCR a cDNA librarythat contained the sequence of interest, and 2) for use as probes toisolate a clone of the full-length coding sequence for PRO353.

[0649] Based on the DNA36363 consensus sequence, forward and reverse PCRprimers were synthesized as follows:

[0650] forward PCR primer 5′-TACAGGCCCAGTCAGGACCAGGGG-3′ (SEQ ID NO: 74)

[0651] reverse PCR primer 5′-CTGAAGAAGTAGAGGCCGGGACG-3′ (SEQ ID NO: 75).

[0652] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the DNA36363 consensus sequence which had the followingnucleotide sequence:

[0653] hybridization probe

[0654] 5′-CCCGGTGCTTGCGCTGCTGTGACCCCGGTACCTCCATGTACCCGG-3′ (SEQ ID NO:76)

[0655] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO353gene using the probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue.

[0656] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO353 [herein designated as DNA41234-1242](SEQ ID NO: 72) and the derived protein sequence for PRO353.

[0657] The entire nucleotide sequence of DNA41234-1242 is shown in FIG.37 (SEQ ID NO: 72). Clone DNA41234-1242 contains a single open readingframe with an apparent translational initiation site at nucleotidepositions 305-307 and ending at the stop codon at nucleotide positions1148-1150 (FIG. 37). The predicted polypeptide precursor is 281 aminoacids long (FIG. 38). Important regions of the amino acid sequenceencoded by PRO353 include the signal peptide, corresponding to aminoacids 1-26, the start of the mature protein at amino acid position 27, apotential N-glycosylation site, corresponding to amino acids 93-98 and aregion which has homology to a 30 kd adipocyte complement-relatedprotein precursor, corresponding to amino acids 99-281. CloneDNA41234-1242 has been deposited with the ATCC and is assigned ATCCdeposit no. 209618.

[0658] Analysis of the amino acid sequence of the full-length PRO353polypeptides suggests that portions of them possess significant homologyto portions of human and murine complement proteins, thereby indicatingthat PRO353 may be a novel complement protein.

Example 21 Isolation of eDNA clones Encoding Human PRO1318

[0659] The cDNA molecule corresponding to DNA73838-1674 as shown in FIG.39 (SEQ ID NO: 77) was obtained from Curagen, Inc.

Example 22 Isolation of cDNA Clones Encoding Human PRO1600

[0660] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequences is herein designated DNA75516. The consensus DNA sequence wasextended using repeated cycles of BLAST and phrap to extend theconsensus sequence as far as possible using the sources of EST sequencesdiscussed above. Based on the DNA75516 consensus sequence,oligonucleotides were synthesized: 1) to identify by PCR a cDNA librarythat contained the sequence of interest, and 2) for use as probes toisolate a clone of the full-length coding sequence for PRO1600.

[0661] Based on the DNA75516 consensus sequence, oligonucleotide probeswere synthesized as follows:

[0662] 5′-AGACATGGCTCAGTCACTGG-3′ (SEQ ID NO: 81)

[0663] 5′-GACCCCTAAAGGGCCATAG-3′ (SEQ ID NO: 82).

[0664] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened with the probesidentified above. RNA for construction of the cDNA libraries wasisolated from human fetal heart tissue.

[0665] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO1600 [herein designated asDNA77503-1686] (SEQ ID NO: 79) and the derived protein sequence forPRO1600.

[0666] The entire nucleotide sequence of DNA77503-1686 is shown in FIG.41 (SEQ ID NO: 79). Clone DNA77503-1686 contains a single open readingframe with an apparent translational initiation site at nucleotidepositions 6-8 and ending at the stop codon at nucleotide positions408-410 (FIG. 41). The predicted polypeptide precursor is 134 aminoacids long (FIG. 42). Important regions of the amino acid sequence ofPRO1600 are shown in FIG. 42. Clone DNA77503-1686 has been depositedwith the ATCC and is assigned ATCC deposit no. 203362.

Example 23 Isolation of cDNA Clones Encoding Human PRO533

[0667] The EST sequence accession number AF007268, a murine fibroblastgrowth factor (FGF-15) was used to search various public EST databases(e.g., GenBank, Dayhoff, etc.). The search was performed using thecomputer program BLAST or BLAST2 [Altschul et al., Methods inEnzymology, 266:460-480 (1996)] as a comparison of the ECD proteinsequences to a 6 frame translation of the EST sequences. The searchresulted in a hit with GenBank EST AA220994, which has been identifiedas stratagene NT2 neuronal precursor 937230.

[0668] Based on the Genbank EST AA220994 sequence, oligonucleotides weresynthesized: 1) to identify by PCR a cDNA library that contained thesequence of interest, and 2) for use as probes to isolate a clone of thefull-length coding sequence. Forward and reverse PCR primers may rangefrom 20 to 30 nucleotides (typically about 24), and are designed to givea PCR product of 100-1000 bp in length. The probe sequences aretypically 40-55 bp (typically about 50) in length. In order to screenseveral libraries for a source of a full-length clone, DNA from thelibraries was screened by PCR amplification, as per Ausubel et al.,Current Protocols in Molecular Biology, with the PCR primer pair. Apositive library was then used to isolate clones encoding the gene ofinterest using the probe oligonucleotide and one of the PCR primers.

[0669] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified below. A positivelibrary was then used to isolate clones encoding the PRO533 gene usingthe probe oligonucleotide and one of the PCR primers.

[0670] RNA for construction of the cDNA libraries was isolated fromhuman fetal retina. The cDNA libraries used to isolated the cDNA cloneswere constructed by standard methods using commercially availablereagents (e.g., Invitrogen, San Diego, Calif.; Clontech, etc.) The cDNAwas primed with oligo dT containing a NotI site, linked with blunt toSalI hemikinased adaptors, cleaved with NotI, sized appropriately by gelelectrophoresis, and cloned in a defined orientation into a suitablecloning vector (such as pRKB or pRKD; pRK5B is a precursor of pRK5D thatdoes not contain the SfiI site; see, Holmes et al., Science,253:1278-1280 (1991)) in the unique XhoI and NotI sites.

[0671] A cDNA clone was sequenced in its entirety. The full lengthnucleotide sequence of PRO533 is shown in FIG. 45 (SEQ ID NO: 85). CloneDNA49435-1219 contains a single open reading frame with an apparenttranslational initiation site at nucleotide positions 459-461 (FIG. 45;SEQ ID NO: 85). The predicted polypeptide precursor is 216 amino acidslong. Clone DNA47412-1219 has been deposited with ATCC and is assignedATCC deposit no. ATCC 209480.

[0672] Based on a BLAST-2 and FastA sequence alignment analysis of thefull-length sequence, PRO533 shows amino acid sequence identity tofibroblast growth factor (53%).

[0673] The oligonucleotide sequences used in the above procedure werethe following:

[0674] FGF15.forward: 5′-ATCCGCCCAGATGGCTACAATGTGTA-3′ (SEQ ID NO: 87);

[0675] FGF15.probe: 5′ -GCCTCCCGGTCTCCCTGAGCAGTGCCAAACAGCGGCAGTGTA-3′(SEQ ID NO :88);

[0676] FGF15.reverse: 5′-CCAGTCCGGTGACAAGCCCAAA-3′ (SEQ ID NO: 89).

Example 24 Isolation of cDNA Clones Encoding Human PRO301

[0677] A consensus DNA sequence designated herein as DNA35936 wasassembled using phrap as described in Example 1 above. Based on thisconsensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence.

[0678] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified below. A positivelibrary was then used to isolate clones encoding the PRO301 gene usingthe probe oligonucleotide and one of the PCR primers.

[0679] RNA for construction of the cDNA libraries was isolated fromhuman fetal kidney.

[0680] A cDNA clone was sequenced in its entirety. The full lengthnucleotide sequence of native sequence PRO301 is shown in FIG. 47 (SEQID NO: 90). Clone DNA40628-1216 contains a single open reading framewith an apparent translational initiation site at nucleotide positions52-54 (FIG. 47; SEQ ID NO: 90). The predicted polypeptide precursor is299 amino acids long with a predicted molecular weight of 32,583 daltonsand pI of 8.29. Clone DNA40628-1216 has been deposited with ATCC and isassigned ATCC deposit No. ATCC 209432.

[0681] Based on a BLAST and FastA sequence alignment analysis of thefull-length sequence, PRO301 shows amino acid sequence identity to A33antigen precursor (30%) and coxsackie and adenovirus receptor protein(29%).

[0682] The oligonucleotide sequences used in the above procedure werethe following:

[0683] OLI2162 (35936.f1) 5′-TCGCGGAGCTGTGTTCTGTTTCCC-3′ (SEQ ID NO: 92)

[0684] OLI2163 (35936.p1)

[0685] 5′-TGATCGCGATGGGGACAAAGGCGCAAGCTCGAGAGGAAACTGTTGTGCCT-3′ (SEQ IDNO: 93)

[0686] OLI2164 (35936.f2)

[0687] 5′-ACACCTGGTTCAAAGATGGG-3′ (SEQ ID NO: 94)

[0688] OLI2165 (35936.r1)

[0689] 5′-TAGGAAGAGTTGCTGAAGGCACGG-3′ (SEQ ID NO: 95)

[0690] OLI2166 (35936.f3)

[0691] 5′-TTGCCTTACTCAGGTGCTAC-3′ (SEQ ID NO: 96)

[0692] OLI2167 (35936.r2)

[0693] 5′-ACTCAGCAGTGGTAGGAAAG-3′ (SEQ ID NO: 97)

Example 25 Isolation of cDNA Clones Encoding Human PRO187

[0694] A proprietary expressed sequence tag (EST) DNA database(LIFESEQ™, Incyte Pharmaceuticals, Palo Alto, Calif.) was searched andan EST (#843193) was identified which showed homology to fibroblastgrowth factor (FGF-8) also known as androgen-induced growth factor. mRNAwas isolated from human fetal lung tissue using reagents and protocolsfrom Invitrogen, San Diego, Calif. (Fast Track 2). The cDNA librariesused to isolate the cDNA clones were constructed by standard methodsusing commercially available reagents (e.g., Invitrogen, San Diego,Calif., Life Technologies, Gaithersburg, Md.). The cDNA was primed witholigo dT containing a NotI site, linked with blunt to SalI hemikinasedadaptors, cleaved with NotI, sized appropriately by gel electrophoresis,and cloned in a defined orientation into the cloning vector pRK5D usingreagents and protocols from Life Technologies, Gaithersburg, Md. (SuperScript Plasmid System). The double-stranded cDNA was sized to greaterthan 1000 bp and the SalI/NotI linkered cDNA was cloned into XhoI/NotIcleaved vector. pRK5D is a cloning vector that has an sp6 transcriptioninitiation site followed by an SfiI restriction enzyme site precedingthe XhoI/NotI cDNA cloning sites.

[0695] Several libraries from various tissue sources were screened byPCR amplification with the following oligonucleotide probes:IN843193.f(OL1315) (SEO ID NO:100) 5′-CAGTACGTGAGGGACCAGGGCGCCATGA-3′IN843193.r (OLI 317) (SEQ ID NO:101) 5′-CCGGTGACCTGCACGTGCTTGCCA-3′

[0696] A positive library was then used to isolate clones encoding thePRO187 gene using one of the above oligonucleotides and the followingoligonucleotide probe: IN843193.p (OLI316) (SEQ ID NO:102)5′-GCGGATCTGCCGCCTGCTCANCTGGTCGGTCATGGCGCCCT-3′

[0697] A cDNA clone was sequenced in entirety. The entire nucleotidesequence of PRO187 (DNA27864-1155) is shown in FIG. 49 (SEQ ID NO: 98).Clone DNA27864-1155 contains a single open reading frame with anapparent translational initiation site at nucleotide position 1 (FIG.49; SEQ ID NO: 98). The predicted polypeptide precursor is 205 aminoacids long. Clone DNA27864-1155 has been deposited with the ATCC(designation: DNA27864-1155) and is assigned ATCC deposit no. ATCC209375.

[0698] Based on a BLAST and FastA sequence alignment analysis (using theALIGN computer program) of the full-length sequence, the PRO187polypeptide shows 74% amino acid sequence identity (Blast score 310) tohuman fibroblast growth factor-8 (androgen-induced growth factor).

Example 26 Isolation of cDNA Clones Encoding Human PRO337

[0699] A cDNA sequence identified in the amylase screen described inExample 2 above is herein designated DNA42301. The DNA42301 sequence wasthen compared to other EST sequences using phrap as described in Example1 above and a consensus sequence designated herein as DNA28761 wasidentified. Based on this consensus sequence, oligonucleotides weresynthesized: 1) to identify by PCR a cDNA library that contained thesequence of interest, and 2) for use as probes to isolate a clone of thefull-length coding sequence. In order to screen several libraries for asource of a full-length clone, DNA from the libraries was screened byPCR amplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO337 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal brain.

[0700] A cDNA clone was sequenced in its entirety. The full lengthnucleotide sequence of DNA43316-1237 is shown in FIG. 51 (SEQ ID NO:103). Clone DNA43316-1237 contains a single open reading frame with anapparent translational initiation site at nucleotide positions 134-136(FIG. 51; SEQ ID NO: 103). The predicted polypeptide precursor is 344amino acids long. Clone DNA43316-1237 has been deposited with ATCC andis assigned ATCC deposit no. 209487

[0701] Based on a BLAST-2 and FastA sequence alignment analysis of thefull-length sequence, PRO337 shows amino acid sequence identity to ratneurotrimin (97%).

Example 27 Isolation of cDNA Clones Encoding Human PRO1411

[0702] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from an Incytedatabase. This EST cluster sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., GenBank) and a proprietary EST DNA database (LIFESEQ®,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify existinghomologies. One or more of the ESTs were derived from a thryroid tissuelibrary. The homology search was performed using the computer programBLAST or BLAST2 (Altshul et al., Methods in Enzymology 266:460-480(1996)). Those comparisons resulting in a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into a consensus DNA sequence with the program phrap (PhilGreen, University of Washington, Seattle, Wash.). The consensus sequenceobtained therefrom is herein designated DNA56013.

[0703] In light of the sequence homology between the DNA56013 sequenceand an EST sequence contained within the Incyte EST 1444225, the cloneincluding this EST was purchased and the cDNA insert was obtained andsequenced. The sequence of this cDNA insert is shown in FIG. 53 and isherein designated as DNA59212-1627.

[0704] The full length clone shown in FIG. 53 contained a single openreading frame with an apparent translational initiation site atnucleotide positions 184-186 and ending at the stop codon found atnucleotide positions 1504-1506 (FIG. 53; SEQ ID NO: 105). The predictedpolypeptide precursor (FIG. 54, SEQ ID NO: 106) is 440 amino acids long.The signal peptide is at about amino acids 1-21, and the cell attachmentsite is at about amino acids 301-303 of SEQ ID NO: 106. PRO1411 has acalculated molecular weight of approximately 42,208 daltons and anestimated pI of approximately 6.36. Clone DNA59212-1627 was depositedwith the ATCC on Sep. 9, 1998 and is assigned ATCC deposit no. 203245.

[0705] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 54 (SEQ ID NO: 106), revealed sequence identitybetween the PRO1411 amino acid sequence and the following Dayhoffsequences (data from database incorporated herein): MTV023_(—)19,P_R05307, P_W26348, P_P82962, AF000949_(—)1, EBN1_EBV, P_R95107,GRP2_PHAVU, P_R81318, and S74439_(—)1.

Example 28 Isolation of cDNA Clones Encoding Human PRO4356

[0706] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequence is designated herein “DNA80200”. Based upon an observedhomology between the DNA80200 consensus sequence and an EST sequencecontained within Merck EST clone 248287, Merck EST clone 248287 waspurchased and its insert obtained and sequenced, thereby providingDNA86576-2595.

[0707] The entire coding sequence of PRO4356 is shown in FIG. 55 (SEQ IDNO: 107). Clone DNA86576-2595 contains a single open reading frame withan apparent translational initiation site at nucleotide positions 55-57,and an apparent stop codon at nucleotide positions 808-810. Thepredicted polypeptide precursor is 251 amino acids long. CloneDNA86576-2595 has been deposited with ATCC and is assigned ATCC depositno. 203868. The full-length PRO4356 protein shown in FIG. 56 has anestimated molecular weight of about 26,935 daltons and a pI of about7.42.

[0708] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 56 (SEQ ID NO: 108), revealed homology betweenthe PRO4356 amino acid sequence and the following Dayhoff sequencesincorporated herein: RNMAGPIAN_(—)1, UPAR_BOVIN, S42152, AF007789_(—)1,UPAR_RAT, UPAR_MOUSE, P_W31165, P_W31168, P_R44423 and P_W26359.

Example 29 Isolation of cDNA Clones Encoding Human PRO246

[0709] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequence is herein designated DNA30955. Based on the DNA30955 consensussequence, oligonucleotides were synthesized: 1) to identify by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence for PRO246.

[0710] A pair of PCR primers (forward and reverse) were synthesized:

[0711] forward PCR primer 5′-AGGGTCTCCAGGAGAAAGACTC-3′ (SEQ ID NO: 111)

[0712] reverse PCR primer 5′-ATTGTGGGCCTTGCAGACATAGAC-3′ (SEQ ID NO:112)

[0713] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA30955 sequence which had the followingnucleotide sequence

[0714] hybridization probe

[0715] 5′ -GGCCAAGCATCAAAACCTTCAGAACTAATGTACTGGTTCCTCCAGCTCC-3 (SEQ IDNO: 113)

[0716] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO246 gene usingthe probe oligonucleotide and one of the PCR primers.

[0717] RNA for construction of the cDNA libraries was isolated fromhuman fetal liver tissue. DNA sequencing of the clones isolated asdescribed above gave the full-length DNA sequence for PRO246 [hereindesignated as DNA35639-1172] (SEQ ID NO: 109) and the derived proteinsequence for PRO246.

[0718] The entire nucleotide sequence of DNA35639-1172 is shown in FIG.57 (SEQ ID NO: 109). Clone DNA35639-1172 contains a single open readingframe with an apparent translational initiation site at nucleotidepositions 126-128 and ending at the stop codon at nucleotide positions1296-1298 (FIG. 57). The predicted polypeptide precursor is 390 aminoacids long (FIG. 58). Clone DNA35639-1172 has been deposited with ATCCand is assigned ATCC deposit no. ATCC 209396.

[0719] Analysis of the amino acid sequence of the full-length PRO246polypeptide suggests that it possess significant homology to the humancell surface protein HCAR, thereby indicating that PRO246 may be a novelcell surface virus receptor.

Example 30 Isolation of cDNA Clones Encoding Human PRO265

[0720] A consensus DNA sequence was assembled relative to other ESTsequences as described in Example 1 above using phrap. This consensussequence is herein designated DNA33679. Based on the DNA33679 consensussequence, oligonucleotides were synthesized: 1) to identify by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence for PRO265.

[0721] PCR primers (two forward and one reverse) were synthesized:

[0722] forward PCR primer A: 5′-CGGTCTACCTGTATGGCAACC-3′ (SEQ ID NO:116);

[0723] forward PCR primer B: 5′-GCAGGACAACCAGATAAACCAC-3′ (SEQ ID NO:117);

[0724] reverse PCR primer 5′-ACGCAGATTTGAGAAGGCTGTC-3′ (SEQ ID NO: 118)

[0725] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA33679 sequence which had the followingnucleotide sequence

[0726] hybridization probe

[0727] 5′-TTCACGGGCTGCTCTTGCCCAGCTCTTGAAGCTTGAAGAGCTGCAC-3′ (SEQ ID NO:119)

[0728] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with PCR primer pairs identified above. A positive librarywas then used to isolate clones encoding the PRO265 gene using the probeoligonucleotide and one of the PCR primers.

[0729] RNA for construction of the cDNA libraries was isolated fromhuman a fetal brain library.

[0730] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO265 [herein designated as DNA36350-1158](SEQ ID NO: 114) and the derived protein sequence for PRO265.

[0731] The entire nucleotide sequence of DNA36350-1158 is shown in FIG.59 (SEQ ID NO: 114). Clone DNA36350-1158 contains a single open readingframe with an apparent translational initiation site at nucleotidepositions 352-354 and ending at the stop codon at positions 2332-2334(FIG. 59). The predicted polypeptide precursor is 660 amino acids long(FIG. 60). Clone DNA36350-1158 has been deposited with ATCC and isassigned ATCC deposit no. ATCC 209378.

[0732] Analysis of the amino acid sequence of the full-length PRO265polypeptide suggests that portions of it possess significant homology tothe fibromodulin and the fibromodulin precursor, thereby indicating thatPRO265 may be a novel member of the leucine rich repeat family,particularly related to fibromodulin.

Example 31 Isolation of cDNA Clones Encoding Human PRO941

[0733] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA35941. Based on the DNA35941consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO941.

[0734] A pair of PCR primers (forward and reverse) were synthesized:

[0735] forward PCR primer 5′-CTTGACTGTCTCTGAATCTGCACCC-3′ (SEQ ID NO:122)

[0736] reverse PCR primer 5′-AAGTGGTGGAAGCCTCCAGTGTGG-3′ (SEQ ID NO:123)

[0737] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA35941 sequence which had the followingnucleotide sequence hybridization probe5′-CCACTACGGTATTAGAGCAAAAGTTAAAAACCATCATGGTTCCTGGAGCAGC-3′ (SEQ IDNO:124)

[0738] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO941 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[0739] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO941 [herein designated as DNA53906-1368](SEQ ID NO: 120) and the derived protein sequence for PRO941.

[0740] The entire nucleotide sequence of DNA53906-1368 is shown in FIG.61 (SEQ ID NO: 120). Clone DNA53906-1368 contains a single open readingframe with an apparent translational initiation site at nucleotidepositions 37-39 and ending at the stop codon at nucleotide positions2353-2355 (FIG. 61). The predicted polypeptide precursor is 772 aminoacids long (FIG. 62). The full-length PRO941 protein shown in FIG. 62has an estimated molecular weight of about 87,002 daltons and a pI ofabout 4.64. Analysis of the full-length PRO941 sequence shown in FIG. 62(SEQ ID NO: 121) evidences the presence of the following: a signalpeptide from about amino acid 1 to about amino acid 21, potentialN-glycosylation sites from about amino acid 57 to about amino acid 60,from about amino acid 74 to about amino acid 77, from about amino acid419 to about amino acid 422, from about amino acid 47 to about a aboutacid 508 to about amino acid 511, from about amino acid 515 to aboutamino acid 518, from about amino acid 516 to about amino acid 519 andfrom about amino acid 534 to about amino acid 537, and cadherinextracellular repeated domain signature sequences from about amino acid136 to about amino acid 146 and from about amino acid 244 to about aminoacid 254. Clone DNA53906-1368 has been deposited with ATCC on Apr. 7,1998 and is assigned ATCC deposit no. 209747.

[0741] Analysis of the amino acid sequence of the full-length PRO941polypeptide suggests that it possesses significant sequence similarityto a cadherin protein, thereby indicating that PRO941 may be a novelcadherin protein family member. More specifically, an analysis of theDayhoff database (version 35.45 SwissProt 35) evidenced significanthomology between the PRO941 amino acid sequence and the followingDayhoff sequences, I50180, CADA_CHICK, I50178, GEN12782, CADC_HUMAN,P_W25637, A38992, P_R49731, D38992 and G02678.

Example 32 Isolation of cDNA Clones Encoding Human PRO10096

[0742] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from the Incytedatabase, designated herein as 5086173H1. This EST cluster sequence wasthen compared to a variety of expressed sequence tag (EST) databaseswhich included public EST databases (e.g., GenBank) and a proprietaryEST DNA database (LIFESEQ®, Incyte Pharmaceuticals, Palo Alto, Calif.)to identify existing homologies. The homology search was performed usingthe computer program BLAST or BLAST2 (Altshul et al., Methods inEnzymology 266:460-480 (1996)). Those comparisons resulting in a BLASTscore of 70 (or in some cases 90) or greater that did not encode knownproteins were clustered and assembled into a consensus DNA sequence withthe program “phrap” (Phil Green, University of Washington, Seattle,Wash.). The consensus sequence obtained therefrom is herein designatedDNA110880.

[0743] In light of an observed sequence homology between the DNA110880sequence and an EST sequence encompassed within clone no. 5088384 fromthe Incyte database, clone no. 5088384 was purchased and the cDNA insertwas obtained and sequenced. It was found herein that that cDNA insertencoded a full-length protein. The sequence of this cDNA insert is shownin FIG. 63 and is herein designated as DNA125185-2506.

[0744] Clone DNA125185-2506 contains a single open reading frame with anapparent translational initiation site at nucleotide positions 58-60 andending at the stop codon at nucleotide positions 595-597 (FIG. 63). Thepredicted polypeptide precursor is 179 amino acids long (FIG. 64). Thefull-length PRO10096 protein shown in FIG. 64 has an estimated molecularweight of about 20,011 daltons and a pI of about 8.10. Analysis of thefull-length PRO10096 sequence shown in FIG. 64 (SEQ ID NO: 126)evidences the presence of a variety of important polypeptide domains asshown in FIG. 64, wherein the locations given for those importantpolypeptide domains are approximate as described above. CloneDNA125185-2506 has been deposited with ATCC on Dec. 7, 1999 and isassigned ATCC deposit no. 1031-PTA.

Example 33 Isolation of cDNA Clones Encoding Human PRO6003

[0745] A cDNA clone (DNA83568-2692) encoding a native human PRO6003polypeptide was identified using a yeast screen, in a human fetal kidneycDNA library that preferentially represents the 5′ ends of the primarycDNA Clones.

[0746] Clone DNA83568-2692 contains a single open reading frame with anapparent translational initiation site at nucleotide positions 638-640and ending at the stop codon at nucleotide positions 2225-2227 (FIG.65). The predicted polypeptide precursor is 529 amino acids long (FIG.66). The full-length PRO6003 protein shown in FIG. 66 has an estimatedmolecular weight of about 59,583 daltons and a pI of about 6.36.Analysis of the full-length PRO6003 sequence shown in FIG. 66 (SEQ IDNO: 128) evidences the presence of a variety of important polypeptidedomains as shown in FIG. 66, wherein the locations given for thoseimportant polypeptide domains are approximate as described above. CloneDNA83568-2692 has been deposited with ATCC on Jul. 20, 1999 and isassigned ATCC Deposit No. 386-PTA.

[0747] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using the ALIGN-2 sequence alignment analysis of the full-lengthsequence shown in FIG. 66 (SEQ ID NO: 128), evidenced sequence identitybetween the PRO6003 amino acid sequence and the following Dayhoffsequences: P_W58986, PTND7_(—)1, YKZ3_YEAST, CEK04B12_(—)1,AB014464_(—)1, PCU07059_(—)1, S31213, CELF25E2_(—)2 AF036408_(—)1, andAB007932_(—)1.

Example 34 Isolation of cDNA Clones Encoding Human PRO6004

[0748] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA85042. Based upon an observedhomology between the DNA85402 consensus sequence and an EST sequencecontained within Incyte EST clone no. 3078492, that clone was purchasedand its insert obtained and sequenced. The sequence of that insert isherein designated as DNA92259 and is shown in FIGS. 67A-B (SEQ ID NO:129).

[0749] Clone DNA92259 contains a single open reading frame with anapparent translational initiation site at nucleotide positions 16-18 andending at the stop codon at nucleotide positions 1078-1080 (FIGS.67A-B). The predicted polypeptide precursor is 354 amino acids long(FIG. 68). The full-length PRO6004 protein shown in FIG. 68 has anestimated molecular weight of about 38,719 daltons and a pI of about6.12. Analysis of the full-length PRO6004 sequence shown in FIG. 68 (SEQID NO: 130) evidences the presence of a variety of important polypeptidedomains as shown in FIG. 68, wherein the locations given for thoseimportant polypeptide domains are approximate as described above.

[0750] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using the ALIGN-2 sequence alignment analysis of the full-lengthsequence shown in FIG. 68 (SEQ ID NO: 130), evidenced sequence identitybetween the PRO6004 amino acid sequence and the following Dayhoffsequences: P_W05152, LAMP_HUMAN, P_W05157, P_W05155, I56551, OPCM_RAT,AMAL_DROME, DMU78177_(—)1, I37246 and NCA1_HUMAN.

Example 35 Isolation of cDNA Clones Encoding Human PRO350

[0751] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA39493. Based on the DNA39493consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO350.

[0752] A pair of PCR primers (forward and reverse) were synthesized:

[0753] forward PCR primer 5′-TCAGGGCTGCCAGGAAGGAAGAGC-3′ (SEQ ID NO:133)

[0754] reverse PCR primer 5′-GAGGAGGAGAAGGTCTTCAGAAGAAG-3′ (SEQ,ID NO:134)

[0755] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA39493 sequence which had the followingnucleotide sequence

[0756] hybridization probe

[0757] 5′-AGAAGTTCCAGTCAGCCCACAAGATGCCATTGTCCCCCGGCCTCC-3′ (SEQ ID NO:135)

[0758] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO350 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue.

[0759] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO350 [herein designated as DNA44175-1314](SEQ ID NO: 131) and the derived protein sequence for PRO350.

[0760] The entire nucleotide sequence of DNA44175-1314 is shown in FIG.69 (SEQ ID NO: 131). Clone DNA44175-1314 contains a single open readingframe with an apparent translational initiation site at nucleotidepositions 356-358 and ending at the stop codon at nucleotide positions821-823 (FIG. 69). The predicted polypeptide precursor is 155 aminoacids long (FIG. 70). The full-length PRO350 protein shown in FIG. 70has an estimated molecular weight of about 17,194 daltons and a pI ofabout 10.44. Analysis of the full-length PRO350 sequence shown in FIG.70 (SEQ ID NO: 132) evidences the presence of a variety of importantpolypeptide domains as shown in FIG. 70.

Example 36 Use of PRO as a hybridization probe

[0761] The following method describes use of a nucleotide sequenceencoding PRO as a hybridization probe.

[0762] DNA comprising the coding sequence of full-length or mature PROas disclosed herein is employed as a probe to screen for homologous DNAs(such as those encoding naturally-occurring variants of PRO) in humantissue cDNA libraries or human tissue genomic libraries.

[0763] Hybridization and washing of filters containing either libraryDNAs is performed under the following high stringency conditions.Hybridization of radiolabeled PRO-derived probe to the filters isperformed in a solution of 50% formamide, 5×SSC, 0.1% SDS, 0.1% sodiumpyrophosphate, 50 mM sodium phosphate, pH 6.8, 2×Denhardt's solution,and 10% dextran sulfate at 42° C. for 20 hours. Washing of the filtersis performed in an aqueous solution of 0.1×SSC and 0.1% SDS at 42° C.

[0764] DNAs having a desired sequence identity with the DNA encodingfull-length native sequence PRO can then be identified using standardtechniques known in the art.

Example 37 Expression of PRO in E. coli

[0765] This example illustrates preparation of an unglycosylated form ofPRO by recombinant expression in E. coli.

[0766] The DNA sequence encoding PRO is initially amplified usingselected PCR primers. The primers should contain restriction enzymesites which correspond to the restriction enzyme sites on the selectedexpression vector. A variety of expression vectors may be employed. Anexample of a suitable vector is pBR322 (derived from E. coli; seeBolivar et al., Gene, 2:95 (1977)) which contains genes for ampicillinand tetracycline resistance. The vector is digested with restrictionenzyme and dephosphorylated. The PCR amplified sequences are thenligated into the vector. The vector will preferably include sequenceswhich encode for an antibiotic resistance gene, a trp promoter, apolyhis leader (including the first six STII codons, polyhis sequence,and enterokinase cleavage site), the PRO coding region, lambdatranscriptional terminator, and an argU gene.

[0767] The ligation mixture is then used to transform a selected E. colistrain using the methods described in Sambrook et al., supra.Transformants are identified by their ability to grow on LB plates andantibiotic resistant colonies are then selected. Plasmid DNA can beisolated and confirmed by restriction analysis and DNA sequencing.

[0768] Selected clones can be grown overnight in liquid culture mediumsuch as LB broth supplemented with antibiotics. The overnight culturemay subsequently be used to inoculate a larger scale culture. The cellsare then grown to a desired optical density, during which the expressionpromoter is turned on.

[0769] After culturing the cells for several more hours, the cells canbe harvested by centrifugation. The cell pellet obtained by thecentrifugation can be solubilized using various agents known in the art,and the solubilized PRO protein can then be purified using a metalchelating column under conditions that allow tight binding of theprotein.

[0770] PRO may be expressed in E. coli in a poly-His tagged form, usingthe following procedure. The DNA encoding PRO is initially amplifiedusing selected PCR primers. The primers will contain restriction enzymesites which correspond to the restriction enzyme sites on the selectedexpression vector, and other useful sequences providing for efficientand reliable translation initiation, rapid purification on a metalchelation column, and proteolytic removal with enterokinase. ThePCR-amplified, poly-His tagged sequences are then ligated into anexpression vector, which is used to transform an E. coli host based onstrain 52 (W3110 fuhA(tonA) Ion galE rpoHts(htpRts) clpP(lacIq).Transformants are first grown in LB containing 50 mg/ml carbenicillin at30° C. with shaking until an O.D.600 of 3-5 is reached. Cultures arethen diluted 50-100 fold into CRAP media (prepared by mixing 3.57 g(NH₄)₂SO₄, 0.71 g sodium citrate•2H2O, 1.07 g KCl, 5.36 g Difco yeastextract, 5.36 g Sheffield hycase SF in 500 mL water, as well as 110 mMMPOS, pH 7.3, 0.55% (w/v) glucose and 7 mM MgSO₄) and grown forapproximately 20-30 hours at 30° C. with shaking. Samples are removed toverify expression by SDS-PAGE analysis, and the bulk culture iscentrifuged to pellet the cells. Cell pellets are frozen untilpurification and refolding.

[0771]E. coli paste from 0.5 to 1 L fermentations (6-10 g pellets) isresuspended in 10 volumes (w/v) in 7 M guanidine, 20 mM Tris, pH 8buffer. Solid sodium sulfite and sodium tetrathionate is added to makefinal concentrations of 0.1M and 0.02 M, respectively, and the solutionis stirred overnight at 4° C. This step results in a denatured proteinwith all cysteine residues blocked by sulfitolization. The solution iscentrifuged at 40,000 rpm in a Beckman Ultracentifuge for 30 min. Thesupernatant is diluted with 3-5 volumes of metal chelate column buffer(6 M guanidine, 20 mM Tris, pH 7.4) and filtered through 0.22 micronfilters to clarify. The clarified extract is loaded onto a 5 ml QiagenNi-NTA metal chelate column equilibrated in the metal chelate columnbuffer. The column is washed with additional buffer containing 50 mMimidazole (Calbiochem, Utrol grade), pH 7.4., The protein is eluted withbuffer containing 250 mM imidazole. Fractions containing the desiredprotein are pooled and stored at 4° C. Protein concentration isestimated by its absorbance at 280 nm using the calculated extinctioncoefficient based on its amino acid sequence.

[0772] The proteins are refolded by diluting the sample slowly intofreshly prepared refolding buffer consisting of: 20 mM Tris, pH 8.6, 0.3M NaCl, 2.5 M urea, 5 mM cysteine, 20 mM glycine and 1 mM EDTA.Refolding volumes are chosen so that the final protein concentration isbetween 50 to 100 micrograms/ml. The refolding solution is stirredgently at 4° C. for 12-36 hours. The refolding reaction is quenched bythe addition of TFA to a final concentration of 0.4% (pH ofapproximately 3). Before further purification of the protein, thesolution is filtered through a 0.22 micron filter and acetonitrile isadded to 2-10% final concentration. The refolded protein ischromatographed on a Poros R1/H reversed phase column using a mobilebuffer of 0.1% TFA with elution with a gradient of acetonitrile from 10to 80%. Aliquots of fractions with A280 absorbance are analyzed on SDSpolyacrylamide gels and fractions containing homogeneous refoldedprotein are pooled. Generally, the properly refolded species of mostproteins are eluted at the lowest concentrations of acetonitrile sincethose species are the most compact with their hydrophobic interiorsshielded from interaction with the reversed phase resin. Aggregatedspecies are usually eluted at higher acetonitrile concentrations. Inaddition to resolving misfolded forms of proteins from the desired form,the reversed phase step also removes endotoxin from the samples.

[0773] Fractions containing the desired folded PRO polypeptide arepooled and the acetonitrile removed using a gentle stream of nitrogendirected at the solution. Proteins are formulated into 20 mM Hepes, pH6.8 with 0.14 M sodium chloride and 4% mannitol by dialysis or by gelfiltration using G25 Superfine (Pharmacia) resins equilibrated in theformulation buffer and sterile filtered.

[0774] Many of the PRO polypeptides disclosed herein were successfullyexpressed as described above.

Example 38 Expression of PRO in mammalian cells

[0775] This example illustrates preparation of a potentiallyglycosylated form of PRO by recombinant expression in mammalian cells.

[0776] The vector, pRK5 (see EP 307,247, published Mar. 15, 1989), isemployed as the expression vector. Optionally, the PRO DNA is ligatedinto pRK5 with selected restriction enzymes to allow insertion of thePRO DNA using ligation methods such as described in Sambrook et al.,supra. The resulting vector is called pRK5-PRO.

[0777] In one embodiment, the selected host cells may be 293 cells.Human 293 cells (ATCC CCL 1573) are grown to confluence in tissueculture plates in medium such as DMEM supplemented with fetal calf serumand optionally, nutrient components and/or antibiotics. About 10 μgpRK5-PRO DNA is mixed with about 1 μg DNA encoding the VA RNA gene[Thimmappaya et al., Cell, 31:543 (1982)] and dissolved in 500 μl of 1mM Tris-HCl, 0.1 mM EDTA, 0.227 M CaCl₂. To this mixture is added,dropwise, 500 μl of 50 mM HEPES (pH 7.35), 280 mM NaCl, 1.5 mM NaPO₄,and a precipitate is allowed to form for 10 minutes at 25° C. Theprecipitate is suspended and added to the 293 cells and allowed tosettle for about four hours at 37° C. The culture medium is aspiratedoff and 2 ml of 20% glycerol in PBS is added for 30 seconds. The 293cells are then washed with serum free medium, fresh medium is added andthe cells are incubated for about 5 days.

[0778] Approximately 24 hours after the transfections, the culturemedium is removed and replaced with culture medium (alone) or culturemedium containing 200 μCi/ml ³⁵S-cysteine and 200 μCi/ml ³⁵S-methionine.After a 12 hour incubation, the conditioned medium is collected,concentrated on a spin filter, and loaded onto a 15% SDS gel. Theprocessed gel may be dried and exposed to film for a selected period oftime to reveal the presence of PRO polypeptide. The cultures containingtransfected cells may undergo further incubation (in serum free medium)and the medium is tested in selected bioassays.

[0779] In an alternative technique, PRO may be introduced into 293 cellstransiently using the dextran sulfate method described by Somparyrac etal., Proc. Natl. Acad. Sci., 12:7575 (1981). 293 cells are grown tomaximal density in a spinner flask and 700 μg pRK5-PRO DNA is added. Thecells are first concentrated from the spinner flask by centrifugationand washed with PBS. The DNA-dextran precipitate is incubated on thecell pellet for four hours. The cells are treated with 20% glycerol for90 seconds, washed with tissue culture medium, and re-introduced intothe spinner flask containing tissue culture medium, 5 μg/ml bovineinsulin and 0.1 μg/ml bovine transferrin. After about four days, theconditioned media is centrifuged and filtered to remove cells anddebris. The sample containing expressed PRO can then be concentrated andpurified by any selected method, such as dialysis and/or columnchromatography.

[0780] In another embodiment, PRO can be expressed in CHO cells. ThepRK5-PRO can be transfected into CHO cells using known reagents such asCaPO₄ or DEAE-dextran. As described above, the cell cultures can beincubated, and the medium replaced with culture medium (alone) or mediumcontaining a radiolabel such as ³⁵S-methionine. After determining thepresence of PRO polypeptide, the culture medium may be replaced withserum free medium. Preferably, the cultures are incubated for about 6days, and then the conditioned medium is harvested. The mediumcontaining the expressed PRO can then be concentrated and purified byany selected method.

[0781] Epitope-tagged PRO may also be expressed in host CHO cells. ThePRO may be subcloned out of the pRK5 vector. The subclone insert canundergo PCR to fuse in frame with a selected epitope tag such as apoly-his tag into a Baculovirus expression vector. The poly-his taggedPRO insert can then be subcloned into a SV40 driven vector containing aselection marker such as DHFR for selection of stable clones. Finally,the CHO cells can be transfected (as described above) with the SV40driven vector. Labeling may be performed, as described above, to verifyexpression. The culture medium containing the expressed poly-His taggedPRO can then be concentrated and purified by any selected method, suchas by Ni²⁺-chelate affinity chromatography.

[0782] PRO may also be expressed in CHO and/or COS cells by a transientexpression procedure or in CHO cells by another stable expressionprocedure.

[0783] Stable expression in CHO cells is performed using the followingprocedure. The proteins are expressed as an IgG construct(immunoadhesin), in which the coding sequences for the soluble forms(e.g. extracellular domains) of the respective proteins are fused to anIgG I constant region sequence containing the hinge, CH2 and CH2 domainsand/or is a poly-His tagged form.

[0784] Following PCR amplification, the respective DNAs are subcloned ina CHO expression vector using standard techniques as described inAusubel et al., Current Protocols of Molecular Biology, Unit 3.16, JohnWiley and Sons (1997). CHO expression vectors are constructed to havecompatible restriction sites 5′ and 3′ of the DNA of interest to allowthe convenient shuttling of cDNA's. The vector used expression in CHOcells is as described in Lucas et al., Nucl. Acids Res. 24:9 (1774-1779(1996), and uses the SV40 early promoter/enhancer to drive expression ofthe cDNA of interest and dihydrofolate reductase (DHFR). DHFR expressionpermits selection for stable maintenance of the plasmid followingtransfection.

[0785] Twelve micrograms of the desired plasmid DNA is introduced intoapproximately 10 million CHO cells using commercially availabletransfection reagents Superfect® (Quiagen), Dosper® or Fugene®(Boehringer Mannheim). The cells are grown as described in Lucas et al.,supra. Approximately 3×10⁻⁷ cells are frozen in an ampule for furthergrowth and production as described below.

[0786] The ampules containing the plasmid DNA are thawed by placementinto water bath and mixed by vortexing. The contents are pipetted into acentrifuge tube containing 10 mLs of media and centrifuged at 1000 rpmfor 5 minutes. The supernatant is aspirated and the cells areresuspended in 10 mL of selective media (0.2 μm filtered PS20 with 5%0.2 μm diafiltered fetal bovine serum). The cells are then aliquotedinto a 100 mL spinner containing 90 mL of selective media. After 1-2days, the cells are transferred into a 250 mL spinner filled with 150 mLselective growth medium and incubated at 37° C. After another 2-3 days,250 mL, 500 and 2000 mL spinners are seeded with 3×10⁵ cells/mL. Thecell media is exchanged with fresh media by centrifugation andresuspension in production medium. Although any suitable CHO media maybe employed, a production medium described in U.S. Pat. No. 5,122,469,issued Jun. 16, 1992 may actually be used. A 3L production spinner isseeded at 1.2×10⁶ cells/mL. On day 0, the cell number pH ie determined.On day 1, the spinner is sampled and sparging with filtered air iscommenced. On day 2, the spinner is sampled, the temperature shifted to33° C., and 30 mL of 500 g/L glucose and 0.6 mL of 10% antifoam (e.g.,35% polydimethylsiloxane emulsion, Dow Corning 365 Medical GradeEmulsion) taken. Throughout the production, the pH is adjusted asnecessary to keep it at around 7.2. After 10 days, or until theviability dropped below 70%, the cell culture is harvested bycentrifugation and filtering through a 0.22 μm filter. The filtrate waseither stored at 4° C. or immediately loaded onto columns forpurification.

[0787] For the poly-His tagged constructs, the proteins are purifiedusing a Ni-NTA column (Qiagen). Before purification, imidazole is addedto the conditioned media to a concentration of 5 mM. The conditionedmedia is pumped onto a 6 ml Ni-NTA column equilibrated in 20 mM Hepes,pH 7.4, buffer containing 0.3 M NaCl and 5 mM imidazole at a flow rateof 4-5 ml/min. at 4° C. After loading, the column is washed withadditional equilibration buffer and the protein eluted withequilibration buffer containing 0.25 M imidazole. The highly purifiedprotein is subsequently desalted into a storage buffer containing 10 mMHepes, 0.14 M NaCl and 4% mannitol, pH 6.8, with a 25 ml G25 Superfine(Pharmacia) column and stored at −80° C. lmmunioadlhesin (Fc-containing)constructs are purified from the conditioned media as follows. Theconditioned medium is pumped onto a 5 ml Protein A column (Pharmacia)which had been equilibrated in 20 mM Na phosphate buffer, pH 6.8. Afterloading, the column is washed extensively with equilibration bufferbefore elution with 100 mM citric acid, pH 3.5. The eluted protein isimmediately neutralized by collecting 1 ml fractions into tubescontaining 275 μL of 1 M Tris buffer, pH 9. The highly purified proteinis subsequently desalted into storage buffer as described above for thepoly-His tagged proteins. The homogeneity is assessed by SDSpolyacrylamide gels and by N-terminal amino acid sequencing by Edmandegradation.

[0788] Many of the PRO polypeptides disclosed herein were successfullyexpressed as described above.

Example 39 Expression of PRO in Yeast

[0789] The following method describes recombinant expression of PRO inyeast.

[0790] First, yeast expression vectors are constructed for intracellularproduction or secretion of PRO from the ADH2/GAPDH promoter. DNAencoding PRO and the promoter is inserted into suitable restrictionenzyme sites in the selected plasmid to direct intracellular expressionof PRO. For secretion, DNA encoding PRO can be cloned into the selectedplasmid, together with DNA encoding the ADH2/GAPDH promoter, a nativePRO signal peptide or other mammalian signal peptide, or, for example, ayeast alpha-factor or invertase secretory signal/leader sequence, andlinker sequences (if needed) for expression of PRO.

[0791] Yeast cells, such as yeast strain ABI 10, can then be transformedwith the expression plasmids described above and cultured in selectedfermentation media. The transformed yeast supernatants can be analyzedby precipitation with 10% trichloroacetic acid and separation bySDS-PAGE, followed by staining of the gels with Coomassie Blue stain.

[0792] Recombinant PRO can subsequently be isolated and purified byremoving the yeast cells from the fermentation medium by centrifugationand then concentrating the medium using selected cartridge filters. Theconcentrate containing PRO may further be purified using selected columnchromatography resins.

[0793] Many of the PRO polypeptides disclosed herein were successfullyexpressed as described above.

Example 40 Expression of PRO in Baculovirus-Infected Insect Cells

[0794] The following method describes recombinant expression of PRO inBaculovirus-infected insect cells.

[0795] The sequence coding for PRO is fused upstream of an epitope tagcontained within a baculovirus expression vector. Such epitope tagsinclude poly-his tags and immunoglobulin tags (like Fe regions of IgG).A variety of plasmids may be employed, including plasmids derived fromcommercially available plasmids such as pVL1393 (Novagen). Briefly, thesequence encoding PRO or the desired portion of the coding sequence ofPRO such as the sequence encoding the extracellular domain of atransmembrane protein or the sequence encoding the mature protein if theprotein is extracellular is amplified by PCR with primers complementaryto the 5′ and 3′ regions. The 5′ primer may incorporate flanking(selected) restriction enzyme sites. The product is then digested withthose selected restriction enzymes and subcloned into the expressionvector.

[0796] Recombinant baculovirus is generated by co-transfecting the aboveplasmid and BaculoGold™ virus DNA (Pharmingen) into Spodopterafrugiperda (“Sf9”) cells (ATCC CRL 1711) using lipofectin (commerciallyavailable from GIBCO-BRL). After 4-5 days of incubation at 28° C., thereleased viruses are harvested and used for further amplifications.Viral infection and protein expression are performed as described byO'Reilley et al., Baculovirus expression vectors: A Laboratory Manual,Oxford: Oxford University Press (1994).

[0797] Expressed poly-his tagged PRO can then be purified, for example,by Ni²⁺-chelate affinity chromatography as follows. Extracts areprepared from recombinant virus-infected Sf9 cells as described byRupert et al., Nature, 362:175-179 (1993). Briefly, Sf9 cells arewashed, resuspended in sonication buffer (25 mL Hepes, pH 7.9; 12.5 mMMgCl₂; 0.1 InM EDTA; 10% glycerol; 0.1% NP-40; 0.4 M KCl), and sonicatedtwice for 20 seconds on ice. The sonicates are cleared bycentrifugation, and the supernatant is diluted 50-fold in loading buffer(50 mM phosphate, 300 mM NaCl, 10% glycerol, pH 7.8) and filteredthrough a 0.45 μm filter. A Ni²⁺-NTA agarose column (commerciallyavailable from Qiagen) is prepared with a bed volume of 5 mL, washedwith 25 mL of water and equilibrated with 25 mL of loading buffer. Thefiltered cell extract is loaded onto the column at 0.5 mL per minute.The column is washed to baseline A₂₈₀ with loading buffer, at whichpoint fraction collection is started. Next, the column is washed with asecondary wash buffer (50 mM phosphate; 300 mM NaCl, 10% glycerol, pH6.0), which elutes nonspecifically bound protein. After reachingA₂₈₀baseline again, the column is developed with a 0 to 500 mM Imidazolegradient in the secondary wash buffer. One mL fractions are collectedand analyzed by SDS-PAGE and silver staining or Western blot withNi²⁺-NTA-conjugated to alkaline phosphatase (Qiagen). Fractionscontaining the eluted His₁₀-tagged PRO are pooled and dialyzed againstloading buffer.

[0798] Alternatively, purification of the IgG tagged (or Fe tagged) PROcan be performed using known chromatography techniques, including forinstance, Protein A or protein G column chromatography.

[0799] Many of the PRO polypeptides disclosed herein were successfullyexpressed as described above.

Example 41 Preparation of Antibodies that Bind PRO

[0800] This example illustrates preparation of monoclonal antibodieswhich can specifically bind PRO.

[0801] Techniques for producing the monoclonal antibodies are known inthe art and are described, for instance, in Goding, supra. Immunogensthat may be employed include purified PRO, fusion proteins containingPRO, and cells expressing recombinant PRO on the cell surface. Selectionof the immunogen can be made by the skilled artisan without undueexperimentation.

[0802] Mice, such as Balb/c, are immunized with the PRO immunogenemulsified in complete Freund's adjuvant and injected subcutaneously orintraperitoneally in an amount from 1-100 micrograms. Alternatively, theimmunogen is emulsified in MPL-TDM adjuvant (Ribi ImmunochemicalResearch, Hamilton, Mont.) and injected into the animal's hind footpads. The immunized mice are then boosted 10 to 12 days later withadditional immunogen emulsified in the selected adjuvant. Thereafter,for several weeks, the mice may also be boosted with additionalimmunization injections. Serum samples may be periodically obtained fromthe mice by retro-orbital bleeding for testing in ELISA assays to detectanti-PRO antibodies.

[0803] After a suitable antibody titer has been detected, the animals“positive” for antibodies can be injected with a final intravenousinjection of PRO. Three to four days later, the mice are sacrificed andthe spleen cells are harvested. The spleen cells are then fused (using35% polyethylene glycol) to a selected murine myeloma cell line such asP3X63AgU. 1, available from ATCC, No. CRL 1597. The fusions generatehybridoma cells which can then be plated in 96 well tissue cultureplates containing HAT (hypoxanthine, aminopterin, and thymidine) mediumto inhibit proliferation of non-fused cells, mycloma hybrids, and spleencell hybrids.

[0804] The hybridoma cells will be screened in an ELISA for reactivityagainst PRO. Determination of “positive” hybridoma cells secreting thedesired monoclonal antibodies against PRO is within the skill in theart.

[0805] The positive hybridoma cells can be injected intraperitoneallyinto syngeneic Balb/c mice to produce ascites containing the anti-PROmonoclonal antibodies. Alternatively, the hybridoma cells can be grownin tissue culture flasks or roller bottles. Purification of themonoclonal antibodies produced in the ascites can be accomplished usingammonium sulfate precipitation, followed by gel exclusionchromatography. Alternatively, affinity chromatography based uponbinding of antibody to protein A or protein G can be employed.

Example 43 Purification of PRO Polypeptides Using Specific Antibodies

[0806] Native or recombinant PRO polypeptides may be purified by avariety of standard techniques in the art of protein purification. Forexample, pro-PRO polypeptide, mature PRO polypeptide, or pre-PROpolypeptide is purified by immunoaffinity chromatography usingantibodies specific for the PRO polypeptide of interest. In general, animmunoaffinity column is constructed by covalently coupling the anti-PROpolypeptide antibody to an activated chromatographic resin.

[0807] Polyclonal immunoglobulin are prepared from immune sera either byprecipitation with ammonium sulfate or by purification on immobilizedProtein A (Pharmacia LKB Biotechnology, Piscataway, N.J.). Likewise,monoclonal antibodies are prepared from mouse ascites fluid by ammoniumsulfate precipitation or chromatography on immobilized Protein A.Partially purified immunoglobulin is covalently attached to achromatographic resin such as CnBr-activated SEPHAROSE™ (Pharmacia LKBBiotechnology). The antibody is coupled to the resin, the resin isblocked, and the derivative resin is washed according to themanufacturer's instructions.

[0808] Such an immunoaffinity column is utilized in the purification ofPRO polypeptide by preparing a fraction from cells containing PROpolypeptide in a soluble form. This preparation is derived bysolubilization of the whole cell or of a subcellular fraction obtainedvia differential centrifugation by the addition of detergent or by othermethods well known in the art. Alternatively, soluble PRO polypeptidecontaining a signal sequence may be secreted in useful quantity into themedium in which the cells are grown.

[0809] A soluble PRO polypeptide-containing preparation is passed overthe immunoaffinity column, and the column is washed under conditionsthat allow the preferential absorbance of PRO polypeptide (e.g., highionic strength buffers in the presence of detergent). Then, the columnis eluted under conditions that disrupt antibody/PRO polypeptide binding(e.g., a low pH buffer such as approximately pH 2-3, or a highconcentration of a chaotrope such as urea or thiocyanate ion), and PROpolypeptide is collected.

Example 44 Drug Screening

[0810] This invention is particularly useful for screening compounds byusing PRO polypeptides or binding fragment thereof in any of a varietyof drug screening techniques. The PRO polypeptide or fragment employedin such a test may either be free in solution, affixed to a solidsupport, borne on a cell surface, or located intracellularly. One methodof drug screening utilizes eukaryotic or prokaryotic host cells whichare stably transformed with recombinant nucleic acids expressing the PROpolypeptide or fragment. Drugs are screened against such transformedcells in competitive binding assays. Such cells, either in viable orfixed form, can be used for standard binding assays. One may measure,for example, the formation of complexes between PRO polypeptide or afragment and the agent being tested. Alternatively, one can examine thediminution in complex formation between the PRO polypeptide and itstarget cell or target receptors caused by the agent being tested.

[0811] Thus, the present invention provides methods of screening fordrugs or any other agents which can affect a PRO polypeptide-associateddisease or disorder. These methods comprise contacting such an agentwith an PRO polypeptide or fragment thereof and assaying (I) for thepresence of a complex between the agent and the PRO polypeptide orfragment, or (ii) for the presence of a complex between the PROpolypeptide or fragment and the cell, by methods well known in the art.In such competitive binding assays, the PRO polypeptide or fragment istypically labeled. After suitable incubation, free PRO polypeptide orfragment is separated from that present in bound form, and the amount offree or uncomplexed label is a measure of the ability of the particularagent to bind to PRO polypeptide or to interfere with the PROpolypeptide/cell complex.

[0812] Another technique for drug screening provides high throughputscreening for compounds having suitable binding affinity to apolypeptide and is described in detail in WO 84/03564, published on Sep.13, 1984. Briefly stated, large numbers of different small peptide testcompounds are synthesized on a solid substrate, such as plastic pins orsome other surface. As applied to a PRO polypeptide, the peptide testcompounds are reacted with PRO polypeptide and washed. Bound PROpolypeptide is detected by methods well known in the art. Purified PROpolypeptide can also be coated directly onto plates for use in theaforementioned drug screening techniques. In addition, non-neutralizingantibodies can be used to capture the peptide and immobilize it on thesolid support.

[0813] This invention also contemplates the use of competitive drugscreening assays in which neutralizing antibodies capable of binding PROpolypeptide specifically compete with a test compound for binding to PROpolypeptide or fragments thereof. In this manner, the antibodies can beused to detect the presence of any peptide which shares one or moreantigenic determinants with PRO polypeptide.

Example 45 Rational Drug Design

[0814] The goal of rational drug design is to produce structural analogsof biologically active polypeptide of interest (i.e., a PRO polypeptide)or of small molecules with which they interact, e.g., agonists,antagonists, or inhibitors. Any of these examples can be used to fashiondrugs which are more active or stable forms of the PRO polypeptide orwhich enhance or interfere with the function of the PRO polypeptide invivo (c.f., Hodgson, Bio/Technology, 9: 19-21 (1991)).

[0815] In one approach, the three-dimensional structure of the PROpolypeptide, or of an PRO polypeptide-inhibitor complex, is determinedby x-ray crystallography, by computer modeling or, most typically, by acombination of the two approaches. Both the shape and charges of the PROpolypeptide must be ascertained to elucidate the structure and todetermine active site(s) of the molecule. Less often, useful informationregarding the structure of the PRO polypeptide may be gained by modelingbased on the structure of homologous proteins. In both cases, relevantstructural information is used to design analogous PRO polypeptide-likemolecules or to identify efficient inhibitors. Useful examples ofrational drug design may include molecules which have improved activityor stability as shown by Braxton and Wells, Biochemistry, 31:7796-7801(1992) or which act as inhibitors, agonists, or antagonists of nativepeptides as shown by Athauda et al., J. Biochem., 113:742-746 (1993).

[0816] It is also possible to isolate a target-specific antibody,selected by functional assay, as described above, and then to solve itscrystal structure. This approach, in principle, yields a pharmacore uponwhich subsequent drug design can be based. It is possible to bypassprotein crystallography altogether by generating anti-idiotypicantibodies (anti-ids) to a functional, pharmacologically activeantibody. As a mirror image of a mirror image, the binding site of theanti-ids would be expected to be an analog of the original receptor. Theanti-id could then be used to identify and isolate peptides from banksof chemically or biologically produced peptides. The isolated peptideswould then act as the pharmacore.

[0817] By virtue of the present invention, sufficient amounts of the PROpolypeptide may be made available to perform such analytical studies asX-ray crystallography. In addition, knowledge of the PRO polypeptideamino acid sequence provided herein will provide guidance to thoseemploying computer modeling techniques in place of or in addition tox-ray crystallography.

Example 46 Mouse Kidney Mesangial Cell Proliferation Assay (Assay 92)

[0818] This assay shows that certain polypeptides of the invention actto induce proliferation of mammalian kidney mesangial cells and,therefore, are useful for treating kidney disorders associated withdecreased mesangial cell function such as Berger disease or othernephropathies associated with Schönlein-Henoch purpura, celiac disease,dermatitis herpetiformis or Crohn disease. The assay is performed asfollows. On day one, mouse kidney mesangial cells are plated on a 96well plate in growth media (3:1 mixture of Dulbecco's modified Eagle'smedium and Ham's F12 medium, 95% fetal bovine serum, 5% supplementedwith 14 mM HEPES) and grown overnight. On day 2, PRO polypeptides arediluted at 2 concentrations(1% and 0.1%) in serum-free medium and addedto the cells. Control samples are serum-free medium alone. On day 4,20μl of the Cell Titer 96 Aqueous one solution reagent (Progema) wasadded to each well and the colormetric reaction was allowed to proceedfor 2 hours. The absorbance (OD) is then measured at 490 nm. A positivein the assay is anything that gives an absorbance reading which is atleast 15% above the control reading.

[0819] The following polypeptides tested positive in this assay:PRO1272.

Example 47 Detection of PRO Polypeptides That Affect Glucose or FFAUptake by Primary Rat Adipocytes (Assay 94)

[0820] This assay is designed to determine whether PRO polypeptides showthe ability to affect glucose or FFA uptake by adipocyte cells. PROpolypeptides testing positive in this assay would be expected to beuseful for the therapeutic treatment of disorders where either thestimulation or inhibition of glucose uptake by adipocytes would bebeneficial including, for example, obesity, diabetes or hyper- orhypo-insulinemia.

[0821] In a 96 well format, PRO polypeptides to be assayed are added toprimary rat adipocytes, and allowed to incubate overnight. Samples aretaken at 4 and 16 hours and assayed for glycerol, glucose and FFAuptake. After the 16 hour incubation, insulin is added to the media andallowed to incubate for 4 hours. At this time, a sample is taken andglycerol, glucose and FFA uptake is measured. Media containing insulinwithout the PRO polypeptide is used as a positive reference control. Asthe PRO polypeptide being tested may either stimulate or inhibit glucoseand FFA uptake, results are scored as positive in the assay if greaterthan 1.5 times or less than 0.5 times the insulin control.

[0822] The following PRO polypeptides tested positive as eitherstimulators or inhibitors of glucose and/or FFA uptake in this assay:PRO196, PRO185, PRO210, PRO215, PRO242, PRO288, PRO1183, PRO1419PRO9940, PRO301, PRO337 and PRO265.

Example 48 Stimulation of Adult Heart Hypertrophy (Assay 2)

[0823] This assay is designed to measure the ability of various PROpolypeptides to stimulate hypertrophy of adult heart. PRO polypeptidestesting positive in this assay would be expected to be useful for thetherapeutic treatment of various cardiac insufficiency disorders.

[0824] Ventricular myocytes freshly isolated from adult (250 g) SpragueDawley rats are plated at 2000 cell/well in 180 μl volume. Cells areisolated and plated on day 1, the PRO polypeptide-containing testsamples or growth medium only (negative control) (20 μl volume) is addedon day 2 and the cells are then fixed and stained on day 5. Afterstaining, cell size is visualized wherein cells showing no growthenhancement as compared to control cells are given a value of 0.0, cellsshowing small to moderate growth enhancement as compared to controlcells are given a value of 1.0 and cells showing large growthenhancement as compared to control cells are given a value of 2.0. Anydegree of growth enhancement as compared to the negative control cellsis considered positive for the assay.

[0825] The following PRO polypeptides tested positive in this assay:PRO301.

Example 49 Inhibition of Vascular Endothelial Growth Factor (VEGF)Stimulated Proliferation of Endothelial Cell Growth (Assay 9)

[0826] The ability of various PRO polypeptides to inhibit VEGFstimulated proliferation of endothelial cells was tested. Polypeptidestesting positive in this assay are useful for inhibiting endothelialcell growth in mammals where such an effect would be beneficial, e.g.,for inhibiting tumor growth.

[0827] Specifically, bovine adrenal cortical capillary endothelial cells(ACE) (from primary culture, maximum of 12-14 passages) were plated in96-well plates at 500 cells/well per 100 microliter. Assay mediaincluded low glucose DMEM, 10% calf serum, 2 mM glutamine, and 1×penicillin/streptomycin/fungizone. Control wells included the following:(1) no ACE cells added; (2) ACE cells alone; (3) ACE cells plus 5 ng/mlFGF; (4) ACE cells plus 3 ng/ml VEGF; (5) ACE cells plus 3 ng/ml VEGFplus 1 ng/ml TGF-beta; and (6) ACE cells plus 3 ng/ml VEGF plus 5 ng/mlLIF. The test samples, poly-his tagged PRO polypeptides (in 100microliter volumes), were then added to the wells (at dilutions of 1%,0.1% and 0.01%, respectively). The cell cultures were incubated for 6-7days at 37° C./5% CO₂. After the incubation, the media in the wells wasaspirated, and the cells were washed 1× with PBS. An acid phosphatasereaction mixture (100 microliter; 0.1 M sodium acetate, pH 5.5, 0.1%Triton X-100, 10 mM p-nitrophenyl phosphate) was then added to eachwell. After a 2 hour incubation at 37° C., the reaction was stopped byaddition of 10 microliters 1N NaOH. Optical density (OD) was measured ona microplate reader at 405 nm.

[0828] The activity of PRO polypeptides was calculated as the percentinhibition of VEGF (3 ng/ml) stimulated proliferation (as determined bymeasuring acid phosphatase activity at OD 405 nm) relative to the cellswithout stimulation. TGF-beta was employed as an activity reference at 1ng/ml, since TGF-beta blocks 70-90% of VEGF-stimulated ACE cellproliferation. The results are indicative of the utility of the PROpolypeptides in cancer therapy and specifically in inhibiting tumorangiogenesis. Numerical values (relative inhibition) are determined bycalculating the percent inhibition of VEGF stimulated proliferation bythe PRO polypeptides relative to cells without stimulation and thendividing that percentage into the percent inhibition obtained by TGF-βat 1 ng/ml which is known to block 70-90% of VEGF stimulated cellproliferation. The results are considered positive if the PROpolypeptide exhibits 30% or greater inhibition of VEGF stimulation ofendothelial cell growth (relative inhibition 30% or greater).

[0829] The following polypeptide tested positive in this assay: PRO301,PRO187 and PRO246.

Example 50 Stimulatory Activity in Mixed Lymphocyte Reaction (MLR) Assay(Assay 24)

[0830] This example shows that certain polypeptides of the invention areactive as a stimulator of the proliferation of stimulated T-lymphocytes.Compounds which stimulate proliferation of lymphocytes are usefultherapeutically where enhancement of an immune response is beneficial. Atherapeutic agent may take the form of antagonists of the polypeptide ofthe invention, for example, murine-human chimeric, humanized or humanantibodies against the polypeptide.

[0831] The basic protocol for this assay is described in CurrentProtocols in Immunology, unit 3.12; edited by J. E. Coligan, A. M.Kruisbeek, D. H. Marglies, E. M. Shevach, W Strober, National Insitutesof Health, Published by John Wiley & Sons, Inc.

[0832] More specifically, in one assay variant, peripheral bloodmononuclear cells (PBMC) are isolated from mammalian individuals, forexample a human volunteer, by leukopheresis (one donor will supplystimulator PBMCs, the other donor will supply responder PBMCs). Ifdesired, the cells are frozen in fetal bovine serum and DMSO afterisolation. Frozen cells may be thawed overnight in assay media (37° C.,5% CO₂) and then washed and resuspended to 3×10⁶ cells/ml of assay media(RPMI; 10% fetal bovine serum, 1% penicillin/streptomycin, 1% glutamine,1% HEPES, 1% non-essential amino acids, 1% pyruvate). The stimulatorPBMCs are prepared by irradiating the cells (about 3000 Rads).

[0833] The assay is prepared by plating in triplicate wells a mixtureof:

[0834] 100:1 of test sample diluted to 1% or to 0.1%,

[0835] 50:1 of irradiated stimulator cells, and

[0836] 50:1 of responder PBMC cells.

[0837] 100 microliters of cell culture media or 100 microliter ofCD4-IgG is used as the control. The wells are then incubated at 37° C.,5% CO₂ for 4 days. On day 5, each well is pulsed with tritiatedthymidine (1.0 mC/Well; Amersham). After 6 hours the cells are washed 3times and then the uptake of the label is evaluated.

[0838] In another variant of this assay, PBMCs are isolated from thespleens of Balb/c mice and C57B6 mice. The cells are teased from freshlyharvested spleens in assay media (RPMI; 10% fetal bovine serum, 1%penicillin/streptomycin, 1% glutamine, 1% HEPES, 1% non-essential aminoacids, 1% pyruvate) and the PBMCs are isolated by overlaying these cellsover Lympholyte M (Organon Telnika), centrifuging at 2000 rpm for 20minutes, collecting and washing the mononuclear cell layer in assaymedia and resuspending the cells to 1×10⁷ cells/ml of assay media. Theassay is then conducted as described above.

[0839] Positive increases over control are considered positive withincreases of greater than or equal to 180% being preferred. However, anyvalue greater than control indicates a stimulatory effect for the testprotein.

[0840] The following PRO polypeptides tested positive in this assay:PRO533 and PRO301.

Example 51 PDB12 Cell Proliferation (Assay 29)

[0841] This example demonstrates that various PRO polypeptides haveefficacy in inducing proliferation of PDB12 pancreatic ductal cells andare, therefore, useful in the therapeutic treatment of disorders whichinvolve protein secretion by the pancreas, including diabetes, and thelike.

[0842] PDB12 pancreatic ductal cells are plated on fibronectin coated 96well plates at 1.5×10³ cells per well in 100, μL/180 μL of growth media.100 μL of growth media with the PRO polypeptide test sample or negativecontrol lacking the PRO polypeptide is then added to well, for a finalvolume of 200 μL. Controls contain growth medium containing a proteinshown to be inactive in this assay. Cells are incubated for 4 days at37° C. 20 μL of Alamar Blue Dye (AB) is then added to each well and theflourescent reading is measured at 4 hours post addition of AB, on amicrotiter plate reader at 530 nm excitation and 590 nm emission. Thestandard employed is cells without Bovine Pituitary Extract (BPE) andwith various concentrations of BPE. Buffer or growth medium onlycontrols from unknowns are run 2 times on each 96 well plate.

[0843] Percent increase in protein production is calculated by comparingthe Alamar Blue Dye calculated protein concentration produced by the PROpolypeptide-treated cells with the Alamar Blue Dye calculated proteinconcentration produced by the negative control cells. A percent increasein protein production of greater than or equal to 25% as compared to thenegative control cells is considered positive.

[0844] The following PRO polypeptides tested positive in this assay:PRO301.

Example 52 Guinea Pig Vascular Leak (Assay 32)

[0845] This assay is designed to determine whether PRO polypeptides ofthe present invention show the ability to induce vascular permeability.Polypeptides testing positive in this assay are expected to be usefulfor the therapeutic treatment of conditions which would benefit fromenhanced vascular permeability including, for example, conditions whichmay benefit from enhanced local immune system cell infiltration.

[0846] Hairless guinea pigs weighing 350 grams or more were anesthetizedwith Ketamine (75-80 mg/kg) and 5 mg/kg Xylazine intramuscularly. Testsamples containing the PRO polypeptide or a physiological buffer withoutthe test polypeptide are injected into skin on the back of the testanimals with 100 μl per injection site intradermally. There wereapproximately 16-24 injection sites per animal. One ml of Evans blue dye(1% in PBS) is then injected intracardially. Skin vascular permeabilityresponses to the compounds (i.e., blemishes at the injection sites ofinjection) are visually scored by measuring the diameter (in mm) ofblue-colored leaks from the site of injection at 1, 6 and 24 hours postadministration of the test materials. The mm diameter of blueness at thesite of injection is observed and recorded as well as the severity ofthe vascular leakage. Blemishes of at least 5 mm in diameter areconsidered positive for the assay when testing purified proteins, beingindicative of the ability to induce vascular leakage or permeability. Aresponse greater than 7 mm diameter is considered positive forconditioned media samples. Human VEGF at 0.1 μg/100 μl is used as apositive control, inducing a response of 4-8 mm diameter.

[0847] The following PRO polypeptides tested positive in this assay:PRO533.

Example 53 Retinal Neuron Survival (Assay 52)

[0848] This example demonstrates that certain PRO polypeptides haveefficacy in enhancing the survival of retinal neuron cells and,therefore, are useful for the therapeutic treatment of retinal disordersor injuries including, for example, treating sight loss in mammals dueto retinitis pigmentosum, AMD, etc.

[0849] Sprague Dawley rat pups at postnatal day 7 (mixed population:glia and retinal neuronal types) are killed by decapitation followingCO₂ anesthesia and the eyes are removed under sterile conditions. Theneural retina is dissected away from the pigment epithelium and otherocular tissue and then dissociated into a single cell suspension using0.25% trypsin in Ca²⁺, Mg²⁺-free PBS. The retinas are incubated at 37°C. for 7-10 minutes after which the trypsin is inactivated by adding 1ml soybean trypsin inhibitor. The cells are plated at 100,000 cells perwell in 96 well plates in DMEM/F12 supplemented with N2 and with orwithout the specific test PRO polypeptide. Cells for all experiments aregrown at 37° C. in a water saturated atmosphere of 5% CO₂. After 2-3days in culture, cells are stained with calcein AM then fixed using 4%paraformaldehyde and stained with DAPI for determination of total cellcount. The total cells (fluorescent) are quantified at 20× objectivemagnification using CCD camera and NIH image software for MacIntosh.Fields in the well are chosen at random.

[0850] The effect of various concentration of PRO polypeptides arereported herein where percent survival is calculated by dividing thetotal number of calcein AM positive cells at 2-3 days in culture by thetotal number of DAPI-labeled cells at 2-3 days in culture. Anythingabove 30% survival is considered positive.

[0851] The following PRO polypeptides tested positive in this assayusing polypeptide concentrations within the range of 0.01% to 1.0% inthe assay: PRO350.

Example 54 Proliferation of Rat Utricular Supporting Cells (Assay 54)

[0852] This assay shows that certain polypeptides of the invention actas potent mitogens for inner ear supporting cells which are auditoryhair cell progenitors and, therefore, are useful for inducing theregeneration of auditory hair cells and treating hearing loss inmammals. The assay is performed as follows. Rat UEC-4 utricularepithelial cells are aliquoted into 96 well plates with a density of3000 cells/well in 200 μl of serum-containing medium at 33° C. The cellsare cultured overnight and are then switched to serum-free medium at 37°C. Various dilutions of PRO polypeptides (or nothing for a control) arethen added to the cultures and the cells are incubated for 24 hours.After the 24 hour incubation, ³H-thymidine (1 μCi/well) is added and thecells are then cultured for an additional 24 hours. The cultures arethen washed to remove unincorporated radiolabel, the cells harvested andCpm per well determined. Cpm of at least 30% or greater in the PROpolypeptide treated cultures as compared to the control cultures isconsidered a positive in the assay.

[0853] The following polypeptides tested positive in this assay: PRO337.

Example 55 Rod Photoreceptor Cell Survival (Assay 56)

[0854] This assay shows that certain polypeptides of the invention actto enhance the survival/proliferation of rod photoreceptor cells and,therefore, are useful for the therapeutic treatment of retinal disordersor injuries including, for example, treating sight loss in mammals dueto retinitis pigmentosum, AMD, etc.

[0855] Sprague Dawley rat pups at 7 day postnatal (mixed population:glia and retinal neuronal cell types) are killed by decapitationfollowing CO₂ anesthesis and the eyes are removed under sterileconditions. The neural retina is dissected away form the pigmentepithelium and other ocular tissue and then dissociated into a singlecell suspension using 0.25% trypsin in Ca²⁺, Mg²⁺-free PBS. The retinasare incubated at 37° C. for 7-10 minutes after which the trypsin isinactivated by adding 1 ml soybean trypsin inhibitor. The cells areplated at 100,000 cells per well in 96 well plates in DMEM/F12supplemented with N₂. Cells for all experiments are grown at 37° C. in awater saturated atmosphere of 5% CO₂. After 2-3 days in culture, cellare fixed using 4% paraformaldehyde, and then stained using CellTrackerGreen CMFDA. Rho 4D2 (ascites or IgG 1:100), a monoclonal antibodydirected towards the visual pigment rhodopsin is used to detect rodphotoreceptor cells by indirect immunofluorescence. The results arecalculated as % survival: total number of calcein—rhodopsin positivecells at 2-3 days in culture, divided by the total number of rhodopsinpositive cells at time 2-3 days in culture. The total cells(fluorescent) are quantified at 20× objective magnification using a CCDcamera and NIH image software for MacIntosh. Fields in the well arechosen at random.

[0856] The following polypeptides tested positive in this assay: PRO350.

Example 56 Skin Vascular Permeability Assay (Assay 64)

[0857] This assay shows that certain polypeptides of the inventionstimulate an immune response and induce inflammation by inducingmononuclear cell, eosinophil and PMN infiltration at the site ofinjection of the animal. Compounds which stimulate an immune responseare useful therapeutically where stimulation of an immune response isbeneficial. This skin vascular permeability assay is conducted asfollows. Hairless guinea pigs weighing 350 grams or more areanesthetized with ketamine (75-80 mg/Kg) and 5 mg/Kg xylazineintramuscularly (IM). A sample of purified polypeptide of the inventionor a conditioned media test sample is injected intradermally onto thebacks of the test animals with 100 μl per injection site. It is possibleto have about 10-30, preferably about 16-24, injection sites per animal.One Al of Evans blue dye (1% in physiologic buffered saline) is injectedintracardially. Blemishes at the injection sites are then measured (mmdiameter) at 1 hr and 6 hr post injection. Animals were sacrificed at 6hrs after injection. Each skin injection site is biopsied and fixed informalin. The skins are then prepared for histopathologic evaluation.Each site is evaluated for inflammatory cell infiltration into the skin.Sites with visible inflammatory cell inflammation are scored aspositive. Inflammatory cells may be neufrophilic, eosinophilic,monocytic or lymphocytic. At least a minimal perivascular infiltrate atthe injection site is scored as positve, no infiltrate at the site ofinjection is scored as negative.

[0858] The following polypeptide tested positive in this assay: PRO301.

Example 57 Induction of Endothelial Cell Apoptosis (Assay 73)

[0859] The ability of PRO polypeptides to induce apoptosis inendothelial cells was tested in human venous umbilical vein endothelialcells (HUVEC, Cell Systems). A positive test in the assay is indicativeof the usefulness of the polypeptide in therapeutically treating tumorsas well as vascular disorders where inducing apoptosis of endothelialcells would be beneficial.

[0860] The cells were plated on 96-well microtiter plates (Amersham LifeScience, cytostar-T scintillating microplate, RPNQI60, sterile,tissue-culture treated, individually wrapped), in 10% serum (CSG-medium,Cell Systems), at a density of 2'10⁴ cells per well in a total volume of100 μl. On day 2, test samples containing the PRO polypeptide were addedin triplicate at dilutions of 1%, 0.33% and 0.11%. Wells without cellswere used as a blank and wells with cells only were used as a negativecontrol. As a positive control 1:3 serial dilutions of 50 μl of a 3×stock of staurosporine were used. The ability of the PRO polypeptide toinduce apoptosis was determined by processing of the 96 well plates fordetection of Annexin V, a member of the calcium and phospholipid bindingproteins, to detect apoptosis. 0.2 ml Annexin V-Biotin stock solution(100 μg/ml) was diluted in 4.6 ml 2×Ca²⁺ binding buffer and 2.5% BSA(1:25 dilution). 50 μl of the diluted Annexin V-Biotin solution wasadded to each well (except controls) to a final concentration of 1.0μg/ml. The samples were incubated for 10-15 minutes with Annexin-Biotinprior to direct addition of ³⁵S-Streptavidin. ³⁵S-Streptavidin wasdiluted in 2×Ca²⁺ Binding buffer BSA and was added to all wells at afinal concentration of 3×10⁴ cpm/well. The plates were then sealed,centrifuged at 1000 rpm for 15 minutes and placed on orbital shaker for2 hours. The analysis was performed on a 1450 Microbeta Trilux (Wallac).Percent above background represents the percentage amount of counts perminute above the negative controls. Percents greater than or equal to30% above background are considered positive.

[0861] The following PRO polypeptides tested positive in this assay:PRO301.

Example 58 Induction of c-fos in Cortical Neurons (Assay 83)

[0862] This assay is designed to determine whether PRO polypeptides showthe ability to induce c-fos in cortical neurons. PRO polypeptidestesting positive in this assay would be expected to be useful for thetherapeutic treatment of nervous system disorders and injuries whereneuronal proliferation would be beneficial.

[0863] Cortical neurons are dissociated and plated in growth medium at10,000 cells per well in 96 well plates. After aproximately 2 cellulardivisions, the cells are treated for 30 minutes with the PRO polypeptideor nothing (negative control). The cells are then fixed for 5 minuteswith cold methanol and stained with an antibody directed againstphosphorylated CREB. mRNA levels are then calculated usingchemiluminiescence. A positive in the assay is any factor that resultsin at least a 2-fold increase in c-fos message as compared to thenegative controls.

[0864] The following PRO polypeptides tested positive in this assay:PRO288.

Example 59 Induction of Pancreatic 13-Cell Precursor Differentiation(Assay 89)

[0865] This assay shows that certain polypeptides of the invention actto induce differentiation of pancreatic β-cell precursor cells intomature pancreatic ,-cells and, therefore, are useful for treatingvarious insulin deficient states in mammals, including diabetesmellitus. The assay is performed as follows. The assay uses a primaryculture of mouse fetal pancreatic cells and the primary readout is analteration in the expression of markers that represent either β-cellprecursors or mature β-cells. Marker expression is measured by real timequantitative PCR (RTQ-PCR); wherein the marker being evaluated isinsulin.

[0866] The pancreata are dissected from E14 embryos (CDI mice). Thepancreata are then digested with collagenase/dispase in F12/DMEM at 37°C. for 40 to 60 minutes (collagenase/dispase, 1.37 mg/ml, BoehringerMannheim, #1097113). The digestion is then neutralized with an equalvolume of 5% BSA and the cells are washed once with RPM! 1640. At day 1,the cells are seeded into 12-well tissue culture plates (pre-coated withlaminin, 20 μg/ml in PBS, Boehringer Mannheim, #124317). Cells frompancreata from 1-2 embryos are distributed per well. The culture mediumfor this primary cuture is 14F/1640. At day 2, the media is removed andthe attached cells washed with RPMI/1640. Two mls of minimal media areadded in addition to the protein to be tested. At day 4, the media isremoved and RNA prepared from the cells and marker expression analyzedby real time quantitative RT-PCR. A protein is considered to be activein the assay if it increases the expression of the relevant β-cellmarker as compared to untreated controls.

[0867] 14F/1640 is RPM11640 (Gibco) plus the following:

[0868] group A 1:1000

[0869] group B 1:1000

[0870] recombinant human insulin 10 μg/ml

[0871] Aprotinin (50 μg/ml) 1:2000 (Boehringer manheim #981532)

[0872] Bovine pituitary extract (BPE) 60 μg/ml

[0873] Gentamycin 100 ng/ml

[0874] Group A: (in 10 ml PBS)

[0875] Transferrin, 100 mg (Sigma T2252)

[0876] Epidermal Growth Factor, 100 μg (BRL 100004)

[0877] Triiodothyronine, 10 μl of 5×10⁻⁶ M (Sigma T5516)

[0878] Ethanolamine, 100 μl of 10⁻¹ M (Sigma E0135)

[0879] Phosphoethalamine, 100 μl of 10⁻¹ M (Sigma P0503)

[0880] Selenium, 4 μl of 10⁻¹ M (Aesar #12574)

[0881] Group C: (in 10 ml 100% ethanol)

[0882] Hydrocortisone, 2 μl of 5×10⁻³ M (Sigma #H0135)

[0883] Progesterone, 100 μl of 1×10⁻³ M (Sigma #P6149)

[0884] Forskolin, 500 μl of 20 mM (Calbiochem #344270)

[0885] Minimal media:

[0886] RPMI 1640 plus transferrin (10 μg/ml), insulin (1 μg/ml),gentamycin (100 ng/ml), aprotinin (50 μg/nml) and BPE (15 μg/ml).

[0887] Defined media:

[0888] RPMI 1640 plus transferrin (10 μg/hl), insulin (1 μg/ml),gentamycin (100 μng/ml) and aprotinin (50 μg/ml).

[0889] The following polypeptides were positive in this assay: PRO1361,PRO1308, PRO1600 and PRO4356.

Example 60 Pericyte c-Fos Induction (Assay 93)

[0890] This assay shows that certain polypeptides of the invention actto induce the expression of c-fos in pericyte cells and, therefore, areuseful not only as diagnostic markers for particular types ofpericyte-associated tumors but also for giving rise to antagonists whichwould be expected to be useful for the therapeutic treatment ofpericyte-associated tumors. Induction of c-fos expression in pericytesis also indicative of the induction of angiogenesis and, as such, PROpolypeptides capable of inducing the expression of c-fos would beexpected to be useful for the treatment of conditions where inducedangiogenesis would be beneficial including, for example, wound healing,and the like. Specifically, on day 1, pericytes are received from VECTechnologies and all but 5 ml of media is removed from flask. On day 2,the pericytes are trypsinized, washed, spun and then plated onto 96 wellplates. On day 7, the media is removed and the pericytes are treatedwith 100 μl of PRO polypeptide test samples and controls (positivecontrol=DME+5% serum+/− PDGF at 500 ng/ml; negative control=protein 32).Replicates are averaged and SD/CV are determined. Fold increase overProtein 32 (buffer control) value indicated by chemiluminescence units(RLU) luminometer reading verses frequency is plotted on a histogram.Two-fold above Protein 32 value is considered positive for the assay.ASY Matrix: Growth media=low glucose DMEM=20% FBS+1×penstrep+1×fungizone. Assay Media=low glucose DMEM+5% FBS.

[0891] The following polypeptides tested positive in this assay: PRO444and PRO217.

Example 61 Detection of Polypeptides That Affect Glucose or FFA Uptakein Skeletal Muscle (Assay 106)

[0892] This assay is designed to determine whether PRO polypeptides showthe ability to affect glucose or FFA uptake by skeletal muscle cells.PRO polypeptides testing positive in this assay would be expected to beuseful for the therapeutic treatment of disorders where either thestimulation or inhibition of glucose uptake by skeletal muscle would bebeneficial including, for example, diabetes or hyper- orhypo-insulinemia.

[0893] In a 96 well format, PRO polypeptides to be assayed are added toprimary rat differentiated skeletal muscle, and allowed to incubateovernight. Then fresh media with the PRO polypeptide and +/− insulin areadded to the wells. The sample media is then monitored to determineglucose and FFA uptake by the skeletal muscle cells. The insulin willstimulate glucose and FFA uptake by the skeletal muscle, and insulin inmedia without the PRO polypeptide is used as a positive control, and alimit for scoring. As the PRO polypeptide being tested may eitherstimulate or inhibit glucose and FFA uptake, results are scored aspositive in the assay if greater than 1.5 times or less than 0.5 timesthe insulin control.

[0894] The following PRO polypeptides tested positive as eitherstimulators or inhibitors of glucose and/or FFA uptake in this assay:PRO196, PRO183, PRO185, PRO215, PRO288, PRO1361, PRO1600, PRO4999,PRO7170, PRO533 and PRO187.

Example 62 Fetal Hemoglobin Induction in an Erythroblastic Cell Line(Assay 107)

[0895] This assay is useful for screening PRO polypeptides for theability to induce the switch from adult hemoglobin to fetal hemoglobinin an erythroblastic cell line. Molecules testing positive in this assayare expected to be useful for therapeutically treating various mammalianhemoglobin-associated disorders such as the various thalasseinias. Theassay is performed as follows. Erythroblastic cells are plated instandard growth medium at 1000 cells/well in a 96 well format. PROpolypeptides are added to the growth medium at a concentration of 0.2%or 2% and the cells are incubated for 5 days at 37° C. As a positivecontrol, cells are treated with 100 μM hemin and as a negative control,the cells arc untreated. After 5 days, cell lysates are prepared andanalyzed for the expression of gamma globin (a fetal marker). A positivein the assay is a gamma globin level at least 2-fold above the negativecontrol.

[0896] The following polypeptides tested positive in this assay:PRO1419.

Example 63 Chondrocyte Re-differentiation Assay (Assay 110)

[0897] This assay shows that certain polypeptides of the invention actto induce redifferentiation of chondrocytes, therefore, are expected tobe useful for the treatment of various bone and/or cartilage disorderssuch as, for example, sports injuries and arthritis. The assay isperformed as follows. Porcine chondrocytes are isolated by overnightcollagenase digestion of articulary cartilage of metacarpophalangealjoints of 4-6 month old female pigs. The isolated cells are then seededat 25,000 cells/cm² in Ham F-12 containing 10% FBS and 4 μg/mlgentamycin. The culture media is changed every third day and the cellsare then seeded in 96 well plates at 5,000 cells/well in 100 μl of thesame media without serum and 100 μl of the test PRO polypeptide, 5 nMstaurosporin (positive control) or medium alone (negative control) isadded to give a final volume of 200 μl/well. After 5 days of incubationat 37° C., a picture of each well is taken and the differentiation stateof the chondrocytes is determined. A positive result in the assay occurswhen the redifferentiation of the chondrocytes is determined to be moresimilar to the positive control than the negative control.

[0898] The following polypeptide tested positive in this assay: PRO215,PRO353, PRO365, PRO1272, PRO301 and PRO337.

Example 64 Chondrocyte Proliferation Assay (Assay 111)

[0899] This assay is designed to determine whether PRO polypeptides ofthe present invention show the ability to induce the proliferationand/or redifferentiation of chondrocytes in culture. PRO polypeptidestesting positive in this assay would be expected to be useful for thetherapeutic treatment of various bone and/or cartilage disorders suchas, for example, sports injuries and arthritis.

[0900] Porcine chondrocytes are isolated by overnight collagenasedigestion of articular cartilage of the metacarpophalangeal joint of 4-6month old female pigs. The isolated cells are then seeded at 25,000cells/cm² in Ham F-12 containing 10% FBS and 4 μg/ml gentamycin. Theculture media is changed every third day and the cells are reseeded to25,000 cells/cm² every five days. On day 12, the cells are seeded in 96well plates at 5,000 cells/well in 100 μl of the same media withoutserum and 100 μl of either serum-free medium (negative control),staurosporin (final concentration of 5 nM; positive control) or the testPRO polypeptide are added to give a final volume of 200 μl/well. After 5days at 37° C., 20 μl of Alamar blue is added to each well and theplates are incubated for an additional 3 hours at 37° C. Thefluorescence is then measured in each well (Ex:530 nm; Em: 590 nm). Thefluorescence of a plate containing 200 μl of the serum-free medium ismeasured to obtain the background. A positive result in the assay isobtained when the fluorescence of the PRO polypeptide treated sample ismore like that of the positive control than the negative control.

[0901] The following PRO polypeptides tested positive in this assay:PRO215, PRO217, PRO248, PRO1361, PRO1419, PRO533 and PRO265.

Example 65 Mouse Mesengial Cell Inhibition Assay (Assay 114)

[0902] This assay is designed to determine whether PRO polypeptides ofthe present invention show the ability to inhibit the proliferation ofmouse mesengial cells in culture. PRO polypeptides testing positive inthis assay would be expected to be useful for the therapeutic treatmentof such diseases or conditions where inhibition of mesengial cellproliferation would be beneficial such as, for example, cystic renaldysplasia, polycystic kidney disease, or other kidney diseaseassoiciated with abnormal mesengial cell proliferation, renal tumors,and the like.

[0903] On day 1, mouse mesengial cells are plated on a 96 well plate ingrowth medium (a 3:1 mixture of Dulbecco's modified Eagle's medium andHam's F 12 medium, 95%; fetal bovine serum, 5%; supplemented with 14mMHEPES) and then are allowed to grow overnight. On day 2, the PROpolypeptide is diluted at 2 different concentrations (1%, 0.1%) inserum-free medium and is added to the cells. The negative control isgrowth medium without added PRO polypeptide. After the cells are allowedto incubate for 48 hours, 20 μl of the Cell Titer 96 Aqueous onesolution reagent (Promega) is added to each well and the colormetricreaction is allowed to proceed for 2 hours. The absorbance (OD) is thenmeasured at 490 nm. A positive in the assay is an absorbance readingwhich is at least 10% above the negative control.

[0904] The following PRO polypeptides tested positive in this assay:PRO1318.

Example 66 Induction of Pancreatic β-Cell Precursor Proliferation (Assay117)

[0905] This assay shows that certain polypeptides of the invention actto induce an increase in the number of pancreatic β-cell precursor cellsand, therefore, are useful for treating various insulin deficient statesin mammals, including diabetes mellitus. The assay is performed asfollows. The assay uses a primary culture of mouse fetal pancreaticcells and the primary readout is an alteration in the expression ofmarkers that represent either β-cell precursors or mature β-cells.Marker expression is measured by real time quantitative PCR (RTQ-PCR);wherein the marker being evaluated is a transcription factor calledPdx1.

[0906] The pancreata are dissected from E14 embryos (CD1 mice). Thepancreata are then digested with collagenase/dispase in F12/DMEM at 37°C. for 40 to 60 minutes (collagenase/dispase, 1.37 mg/ml, BoehringerMannheim, #1097113). The digestion is then neutralized with an equalvolume of 5% BSA and the cells are washed once with RPMI1640. At day 1,the cells are seeded into 12-well tissue culture plates (pre-coated withlaminin, 20 μg/mi in PBS, Boehringer Mannheim, #124317). Cells frompancreata from 1-2 embryos are distributed per well. The culture mediumfor this primary cuture is 14F/1640. At day 2, the media is removed andthe attached cells washed with RPMI/1640. Two mls of minimal media areadded in addition to the protein to be tested. At day 4, the media isremoved and RNA prepared from the cells and marker expression analyzedby real time quantitative RT-PCR. A protein is considered to be activein the assay if it increases the expression of the relevant β-cellmarker as compared to untreated controls.

[0907] 14F/1640 is RPMI1640 (Gibco) plus the following:

[0908] group A 1:1000

[0909] group B 1:1000

[0910] recombinant human insulin 10 μg/ml

[0911] Aprotinin (50 μg/mnl) 1:2000 (Boehringer manheim #981532)

[0912] Bovine pituitary extract (BPE) 60 μg/ml

[0913] Gentamycin 100 ng/ml

[0914] Group A: (in 10 ml PBS)

[0915] Transferrin, 100 mg (Sigma T2252)

[0916] Epidermal Growth Factor, 100 μg (BRL 100004)

[0917] Triiodothyronine,10 μl of 5×10⁻⁶ M (Sigma T5516)

[0918] Ethanolamine, 100 μl of 10⁻¹ M (Sigma E0135)

[0919] Phosphoethalamine, 100 μl of 10⁻¹ M (Sigma P0503)

[0920] Selenium, 4 μl of 10⁻¹ M (Aesar #12574)

[0921] Group C: (in 10 ml 100% ethanol)

[0922] Hydrocortisone, 2 μl of 5×10⁻³ M (Sigma #H0135)

[0923] Progesterone, 100 μl of 1×10⁻³ M (Sigma #P6149)

[0924] Forskolin, 500 μl of 20 mM (Calbiochem #344270)

[0925] Minimal media:

[0926] RPMI 1640 plus transferrin (10 μg/ml), insulin (1 μg/ml),gentamycin (100 ng/ml), aprotinin (50 μg/ml) and BPE (15 μg/ml).

[0927] Defined media:

[0928] RPMI 1640 plus transferrin (10 μg/ml), insulin (1 μg/ml),gentamycin (100 ng/ml) and aprotinin (50 μg/ml).

[0929] The following polypeptides tested positive in this assay: PRO183,PRO185, PRO288.

Example 67 In Vitro Antitumor Assay (Assay 161)

[0930] The antiproliferative activity of various PRO polypeptides wasdetermined in the investigational, disease-oriented in vitro anti-cancerdrug discovery assay of the National Cancer Institute (NCI), using asulforhodamine B (SRB) dye binding assay essentially as described bySkehan et al., J. Natl. Cancer Inst. 82:1107-1112 (1990). The 60 tumorcell lines employed in this study (“the NCI panel”), as well asconditions for their maintenance and culture in vitro have beendescribed by Monks et al., J. Natl. Cancer Inst. 83:757-766(1991). Thepurpose of this screen is to initially evaluate the cytotoxic and/orcytostatic activity of the test compounds against different types oftumors (Monks et al., supra; Boyd, Cancer: Princ. Pract. Oncol. Update3(10):1-12 [1989]).

[0931] Cells from approximately 60 human tumor cell lines were harvestedwith trypsin/EDTA (Gibco), washed once, resuspended in IMEM and theirviability was determined. The cell suspensions were added by pipet (100μL volume) into separate 96-well microtiter plates. The cell density forthe 6-day incubation was less than for the 2-day incubation to preventovergrowth. Inoculates were allowed a preincubation period of 24 hoursat 37° C. for stabilization. Dilutions at twice the intended testconcentration were added at time zero in 100 μL aliquots to themicrotiter plate wells (1:2 dilution). Test compounds were evaluated atfive half-log dilutions (1000 to 100,000-fold). Incubations took placefor two days and six days in a 5% CO₂ atmosphere and 100% humidity.

[0932] After incubation, the medium was removed and the cells were fixedin 0.1 ml of 10% trichloroacetic acid at 40° C. The plates were rinsedfive times with deionized water, dried, stained for 30 minutes with 0.1ml of 0.4% sulforhodamine B dye (Sigma) dissolved in 1% acetic acid,rinsed four times with 1% acetic acid to remove unbound dye, dried, andthe stain was extracted for five minutes with 0.1 ml of 10 mM Tris base[tris(hydroxymethyl)aminomethane], pH 10.5. The absorbance (OD) ofsulforhodamine B at 492 nm was measured using a computer-interfaced,96-well microtiter plate reader.

[0933] A test sample is considered positive if it shows at least 50%growth inhibitory effect at one or more concentrations. The positiveresults are shown in the following Table 7. TABLE 7 Compound Tumor TypeDesignation PRO301 NSCL NCI-H322M PRO301 Leukemia MOLT-4; SR PRO301 NSCLA549/ATCC; EKVX; PRO301 NSCL NCI-H23; NCI-460; NCI-H226 PRO301 ColonCOLO 205; HCC-2998; PRO301 Colon HCT-15; KM12; HT29; PRO301 ColonHCT-116 PRO301 CNS SF-268; SF-295; SNB-19 PRO301 Melanoma MALME-3M;SK-MEL-2; PRO301 Melanoma SK-MEL-5; UACC-257 PRO301 Melanoma UACC-62PRO301 Ovarian IGROV1; OVCAR-4 PRO301 Ovarian OVCAR-5 PRO301 OvarianOVCAR-8; SK0OV-3 PRO301 Renal ACHN; CAKI-1; TK-10; UO-31 PRO301 ProstatePC-3; DU-145 PRO301 Breast NCI/ADR-RES; HS 578T PRO301 BreastMDA-MB-435; MDA-N; T-47D PRO301 Melanoma M14 PRO301 Leukemia CCRF-CEM;HL-60(TB); K-562 PRO301 Leukemia RPMI-8226 PRO301 Melanoma LOX IMVIPRO301 Renal 786-0; SN12C PRO301 Breast MCF7; MDA-MB-231/ATCC PRO301Breast BT-549 PRO301 NSCL HOP-62 PRO301 CNS SF-539 PRO301 OvarianOVCAR-3

[0934] The results of these assays demonstrate that the positive testingPRO polypeptides are useful for inhibiting neoplastic growth in a numberof different tumor cell types and may be used therapeutically therefor.Antibodies against these PRO polypeptides are useful for affinitypurification of these useful polypeptides. Nucleic acids encoding thesePRO polypeptides are useful for the recombinant preparation of thesepolypeptides.

Example 68 Gene Amplification in Tumors

[0935] This example shows that certain PRO polypeptide-encoding genesare amplified in the genome of certain human lung, colon and/or breastcancers and/or cell lines. Amplification is associated withoverexpression of the gene product, indicating that the polypeptides areuseful targets for therapeutic intervention in certain cancers such ascolon, lung, breast and other cancers and diagnostic determination ofthe presence of those cancers. Therapeutic agents may take the form ofantagonists of the PRO polypeptide, for example, murine-human chimeric,humanized or human antibodies against a PRO polypeptide.

[0936] The starting material for the screen was genomic DNA isolatedfrom a variety cancers. The DNA is quantitated precisely, e.g.,fluorometrically. As a negative control, DNA was isolated from the cellsof ten normal healthy individuals which was pooled and used as assaycontrols for the gene copy in healthy individuals (not shown). The 5′nuclease assay (for example, TaqMan™) and real-time quantitative PCR(for example, ABI Prizm 7700 Sequence Detection System™ (Perkin Elmer,Applied Biosystems Division, Foster City, Calif.)), were used to findgenes potentially amplified in certain cancers. The results were used todetermine whether the DNA encoding the PRO polypeptide isover-represented in any of the primary lung or colon cancers or cancercell lines or breast cancer cell lines that were screened. The primarylung cancers were obtained from individuals with tumors of the type andstage as indicated in Table 8. An explanation of the abbreviations usedfor the designation of the primary tumors listed in Table 8 and theprimary tumors and cell lines referred to throughout this example aregiven below.

[0937] The results of the TaqMan™ are reported in delta (Δ) Ct units.One unit corresponds to 1 PCR cycle or approximately a 2-foldamplification relative to normal, two units corresponds to 4-fold, 3units to 8-fold amplification and so on. Quantitation was obtained usingprimers and a TaqMan™ fluorescent probe derived from the PROpolypeptide-encoding gene. Regions of the PRO polypeptide-encoding genewhich are most likely to contain unique nucleic acid sequences and whichare least likely to have spliced out introns are preferred for theprimer and probe derivation, e.g., 3′-untranslated regions. Thesequences for the primers and probes (forward, reverse and probe) usedfor the PRO polypeptide gene amplification analysis were as follows:

[0938] PRO533 (DNA49435-1219)

[0939] forward: 5′-GGGACGTGCTTCTACAAGAACAG-3′ (SEQ ID NO: 140)

[0940] reverse: 5′-CAGGCTTACAATGTTATGATCAGACA-3′ (SEQ ID NO: 141)

[0941] probe: 5′-TATTCAGAGTTTTCCATTGGCAGTGCCAGTT-3′ (SEQ ID NO: 142)

[0942] PRO187 (DNA27864-1155)

[0943] forward: 5′-GGCCTTGCAGACAACCGT-3′ (SEQ ID NO: 143)

[0944] reverse: 5′-CAGACTGAGGGAGATCCGAGA-3′ (SEQ ID NO: 144)

[0945] probe: 5′-GCAGATTTTGAGGACAGCCACCTCCA-3′ (SEQ ID NO: 145)

[0946] forward2: 5′-CATCAAGCGCCTCTACCA-3′ (SEQ ID NO: 146)

[0947] reverse2: 5′-CACAAACTCGAACTGCTTCTG-3′ (SEQ ID NO: 147)

[0948] probe2: 5′-CAGCTGCCCTTCCCCAACCA-3′ (SEQ ID NO: 148)

[0949] PRO246 (DNA35639-1172)

[0950] forward: 5′-GGCAGAGACTTCCAGTCACTGA-3′ (SEQ ID NO: 149)

[0951] reverse: 5′-GCCAAGGGTGGTGTTAGATAGG-3′ (SEQ ID NO: 150)

[0952] probe: 5′-CAGGCCCCCTTGATCTGTACCCCA-3′ (SEQ ID NO: 151)

[0953] The 5′ nuclease assay reaction is a fluorescent PCR-basedtechnique which makes use of the 5′ exonuclease activity of Taq DNApolymerase enzyme to monitor amplification in real time. Twooligonucleotide primers (forward [.f] and reverse [.r]) are used togenerate an amplicon typical of a PCR reaction. A third oligonucleotide,or probe (.p), is designed to detect nucleotide sequence located betweenthe two PCR primers. The probe is non-extendible by Taq DNA polymeraseenzyme, and is labeled with a reporter fluorescent dye and a quencherfluorescent dye. Any laser-induced emission from the reporter dye isquenched by the quenching dye when the two dyes are located closetogether as they are on the probe. During the amplification reaction,the Taq DNA polymerase enzyme cleaves the probe in a template-dependentmanner. The resultant probe fragments disassociate in solution, andsignal from the released reporter dye is free from the quenching effectof the second fluorophore. One molecule of reporter dye is liberated foreach new molecule synthesized, and detection of the unquenched reporterdye provides the basis for quantitative interpretation of the data.

[0954] The 5′ nuclease procedure is run on a real-time quantitative PCRdevice such as the ABI Prism 7700TM Sequence Detection. The systemconsists of a thermocycler, laser, charge-coupled device (CCD) cameraand computer. The system amplifies samples in a 96-well format on athermocycler. During amplification, laser-induced fluorescent signal iscollected in real-time through fiber optics cables for all 96 wells, anddetected at the CCD. The system includes software for running theinstrument and for analyzing the data.

[0955] 5′ Nuclease assay data are initially expressed as Ct, or thethreshold cycle. This is defined as the cycle at which the reportersignal accumulates above the background level of fluorescence. The ΔCtvalues are used as quantitative measurement of the relative number ofstarting copies of a particular target sequence in a nucleic acid samplewhen comparing cancer DNA results to normal human DNA results.

[0956] Table 8 describes the stage, T stage and N stage of variousprimary tumors which were used to screen the PRO polypeptide compoundsof the invention. TABLE 8 Primary Lung and Colon Tumor Profiles PrimaryTumor Stage Stage Other Stage Dukes Stage T Stage N Stage Human lungtumor AdenoCa (SRCC724) [LT1] IIA T1 N1 Human lung tumor SqCCa (SRCC725)[LT1a] IIB T3 N0 Human lung tumor AdenoCa (SRCC726) [LT2] IB T2 N0 Humanlung tumor AdenoCa (SRCC727) [LT3] IIIA T1 N2 Human lung tumor AdenoCa(SRCC728) [LT4] IB T2 N0 Human lung tumor SqCCa (SRCC729) [LT6] IB T2 N0Human lung tumor Aden/SqCCa (SRCC730) [LT7] IA T1 N0 Human lung tumorAdenoCa (SRCC731) [LT9] IB T2 N0 Human lung tumor SqCCa (SRCC732) [LT10]IIB T2 N1 Human lung tumor SqCCa (SRCC733) [LT11] IIA T1 N1 Human lungtumor AdenoCa (SRCC734) [LT12] IV T2 N0 Human lung tumor AdenoSqCCa(SRCC735)[LT13] IB T2 N0 Human lung tumor SqCCa (SRCC736) [LT15] IB T2N0 Human lung tumor SqCCa (SRCC737) [LT16] IB T2 N0 Human lung tumorSqCCa (SRCC738) [LT17] IIB T2 N1 Human lung tumor SqCCa (SRCC739) [LT18]IB T2 N0 Human lung tumor SqCCa (SRCC740) [LT19] IB T2 N0 Human lungtumor LCCa (SRCC741) [LT21] IIB T3 N1 Human lung AdenoCa (SRCC811)[LT22] 1A T1 N0 Human colon AdenoCa (SRCC742) [CT2] M1 D pT4 N0 Humancolon AdenoCa (SRCC743) [CT3] B pT3 N0 Human colon AdenoCa (SRCC744)[CT8] B T3 N0 Human colon AdenoCa (SRCC745) [CT10] A pT2 N0 Human colonAdenoCa (SRCC746) [CT12] MO, R1 B T3 N0 Human colon AdenoCa (SRCC747)[CT14] pMO, RO B pT3 pN0 Human colon AdenoCa (SRCC748) [CT15] M1, R2 DT4 N2 Human colon AdenoCa (SRCC749) [CT16] pMO B pT3 pN0 Human colonAdenoCa (SRCC750) [CT17] C1 pT3 pN1 Human colon AdenoCa (SRCC751) [CT1]MO, R1 B pT3 N0 Human colon AdenoCa (SRCC752) [CT4] B pT3 M0 Human colonAdenoCa (SRCC753) [CT5] G2 C1 pT3 pN0 Human colon AdenoCa (SRCC754)[CT6] pMO, RO B pT3 pN0 Human colon AdenoCa (SRCC755) [CT7] G1 A pT2 pN0Human colon AdenoCa (SRCC756) [CT9] G3 D pT4 pN2 Human colon AdenoCa(SRCC757) [CT11] B T3 N0 Human colon AdenoCa (SRCC758) [CT18] MO, RO BpT3 pN0

[0957] DNA Preparation:

[0958] DNA was prepared from cultured cell lines, primary tumors, normalhuman blood. The isolation was performed using purification kit, bufferset and protease and all from Quiagen, according to the manufacturer'sinstructions and the description below.

[0959] Cell culture lysis:

[0960] Cells were washed and trypsinized at a concentration of 7.5×10⁸per tip and pelleted by centrifuging at 1000 rpm for 5 minutes at 4° C.,followed by washing again with ½ volume of PBS recentrifugation. Thepellets were washed a third time, the suspended cells collected andwashed 2× with PBS. The cells were then suspended into 10 ml PBS. BufferC1 was equilibrated at 4° C. Qiagen protease #19155 was diluted into 6.2ml cold ddH₂O to a final concentration of 20 mg/ml and equilibrated at4° C. 10 ml of G2 Buffer was prepared by diluting Qiagen RNAse A stock(100 mg/ml) to a final concentration of 200 μg/ml.

[0961] Buffer C1 (10 ml, 4° C.) and ddH₂O (40 ml, 4° C.) were then addedto the 10 ml of cell suspension, mixed by inverting and incubated on icefor 10 minutes. The cell nuclei were pelleted by centrifuging in aBeckman swinging bucket rotor at 2500 rpm at 4° C. for 15 minutes. Thesupernatant was discarded and the nuclei were suspended with a vortexinto 2 ml Buffer C1 (at 4° C.) and 6 ml ddH₂O, followed by a second 4°C. centrifugation at 2500 rpm for 15 minutes. The nuclei were thenresuspended into the residual buffer using 200 μl per tip. G2 buffer (10ml) was added to the suspended nuclei while gentle vortexing wasapplied. Upon completion of buffer addition, vigorous vortexing wasapplied for 30 seconds. Quiagen protease (200 μl, prepared as indicatedabove) was added and incubated at 50° C. for 60 minutes. The incubationand centrifugation was repeated until the lysates were clear (e.g.,incubating additional 30-60 minutes, pelleting at 3000×g for 10 min., 4°C.).

[0962] Solid human tumor sample preparation and lysis:

[0963] Tumor samples were weighed and placed into 50 ml conical tubesand held on ice. Processing was limited to no more than 250 mg tissueper preparation (1 tip/preparation). The protease solution was freshlyprepared by diluting into 6.25 ml cold ddH₂O to a final concentration of20 mg/ml and stored at 4° C. G2 buffer (20 ml) was prepared by dilutingDNAse A to a final concentration of 200 mg/ml (from 100 mg/ml stock).The tumor tissue was homogenated in 19 ml G2 buffer for 60 seconds usingthe large tip of the polytron in a laminar-flow TC hood in order toavoid inhalation of aerosols, and held at room temperature. Betweensamples, the polytron was cleaned by spinning at 2×30 seconds each in 2LddH₂O, followed by G2 buffer (50 ml). If tissue was still present on thegenerator tip, the apparatus was disassembled and cleaned.

[0964] Quiagen protease (prepared as indicated above, 1.0 ml) was added,followed by vortexing and incubation at 50° C. for 3 hours. Theincubation and centrifugation was repeated until the lysates were clear(e.g., incubating additional 30-60 minutes, pelleting at 3000×g for 10min., 4° C.).

[0965] Human blood preparation and lysis:

[0966] Blood was drawn from healthy volunteers using standard infectiousagent protocols and citrated into 10 ml samples per tip. Quiagenprotease was freshly prepared by dilution into 6.25 ml cold ddH₂O to afinal concentration of 20 mg/ml and stored at 4° C. G2 buffer wasprepared by diluting RNAse A to a final concentration of 200 μg/ml from100 mg/ml stock. The blood (10 ml) was placed into a 50 ml conical tubeand 10 ml C1 buffer and 30 ml ddH₂O (both previously equilibrated to 4°C.) were added, and the components mixed by inverting and held on icefor 10 minutes. The nuclei were pelleted with a Beckman swinging bucketrotor at 2500 rpm, 4° C. for 15 minutes and the supernatant discarded.With a vortex, the nuclei were suspended into 2 ml C1 buffer (4° C) and6 ml ddH₂O (4° C). Vortexing was repeated until the pellet was white.The nuclei were then suspended into the residual buffer using a 200 μltip. G2 buffer (10 ml) were added to the suspended nuclei while gentlyvortexing, followed by vigorous vortexing for 30 seconds. Quiagenprotease was added (200 μl) and incubated at 50° C. for 60 minutes. Theincubation and centrifugation was repeated until the lysates were clear(e.g., incubating additional 30-60 minutes, pelleting at 3000 × g for 10min., 4° C.).

[0967] Purification of cleared lysates:

[0968] (1) Isolation of genomic DNA:

[0969] Genomic DNA was equilibrated (1 sample per maxi tip preparation)with 10 ml QBT buffer. QF elution buffer was equilibrated at 50° C. Thesamples were vortexed for 30 seconds, then loaded onto equilibrated tipsand drained by gravity. The tips were washed with 2'15 ml QC buffer. TheDNA was eluted into 30 ml silanized, autoclaved 30 ml Corex tubes with15 ml QF buffer (50° C.). Isopropanol (10.5 ml) was added to eachsample, the tubes covered with parafin and mixed by repeated inversionuntil the DNA precipitated. Samples were pelleted by centrifugation inthe SS-34 rotor at 15,000 rpm for 10 minutes at 4° C. The pelletlocation was marked, the supernatant discarded, and 10 ml 70% ethanol(4° C.) was added. Samples were pelleted again by centrifugation on theSS-34 rotor at 10,000 rpm for 10 minutes at 4° C. The pellet locationwas marked and the supernatant discarded. The tubes were then placed ontheir side in a drying rack and dried 10 minutes at 3720 C., taking carenot to overdry the samples.

[0970] After drying, the pellets were dissolved into 1.0 ml TE (pH 8.5)and placed at 50° C. for 1-2 hours. Samples were held overnight at 4° C.as dissolution continued. The DNA solution was then transferred to 1.5ml tubes with a 26 gauge needle on a tuberculin syringe. The transferwas repeated 5× in order to shear the DNA. Samples were then placed at50° C. for 1-2 hours.

[0971] (2) Ouantitation of zenomic DNA and preparation for geneamplification assay:

[0972] The DNA levels in each tube were quantified by standard A₂₆₀,A₂₈₀, spectrophotometry on a 1:20 dilution (5 μl DNA+95 μl ddH₂O) usingthe 0.1 ml quartz cuvetts in the Beckman DU640 spectrophotometer.A₂₆₀/A₂₈₀ ratios were in the range of 1.8-1.9. Each DNA samples was thendiluted further to approximately 200 ng/ml in TE (pH 8.5). If theoriginal material was highly concentrated (about 700 ng/μl), thematerial was placed at 50° C. for several hours until resuspended.

[0973] Fluorometric DNA quantitation was then performed on the dilutedmaterial (20-600 ng/ml) using the manufacturer's guidelines as modifiedbelow. This was accomplished by allowing a Hoeffer DyNA Quant 200fluorometer to warm-up for about 15 minutes. The Hoechst dye workingsolution (#H33258, 10 μl, prepared within 12 hours of use) was dilutedinto 100 ml 1× TNE buffer. A 2 ml cuvette was filled with thefluorometer solution, placed into the machine, and the machine waszeroed. pGEM 3Zf(+) (2 μl, lot #360851026) was added to 2 ml offluorometer solution and calibrated at 200 units. An additional 2 μl ofpGEM 3Zf(+) DNA was then tested and the reading confirmed at 400 +/− 10units. Each sample was then read at least in triplicate. When 3 sampleswere found to be within 10% of each other, their average was taken andthis value was used as the quantification value.

[0974] The fluorometricly determined concentration was then used todilute each sample to 10 ng/μl in ddH₂O. This was done simultaneously onall template samples for a single TaqMan plate assay, and with enoughmaterial to run 500-1000 assays. The samples were tested in triplicatewith Taqman™ primers and probe both B-actin and GAPDH on a single platewith normal human DNA and no-template controls. The diluted samples wereused provided that the CT value of normal human DNA subtracted from testDNA was +/− 1 Ct. The diluted, lot-qualified genomic DNA was stored in1.0 ml aliquots at −80° C. Aliquots which were subsequently to be usedin the gene amplification assay were stored at 4° C. Each 1 ml aliquotis enough for 8-9 plates or 64 tests.

[0975] Gene amplification assay:

[0976] The PRO polypeptide compounds of the invention were screened inthe following primary tumors and the resulting ΔCt values greater thanor equal to 1.0 are reported in Table 9 below. TABLE 9 ΔCt values inlung and colon primary tumors and cell line models Primary Tumors orCell Lines PRO187 PRO533 PRO246 LT7 1.04 LT13 2.74 1.63 2.98 1.68 2.44LT3 1.06 LT12 2.70 2.47 2.90 1.74 2.27 LT30 1.67 LT21 1.50 LT-1a 1.02LT10 1.07 LT11 1.09 3.43 1.41 LT15 3.75 2.11 3.92 1.56 3.49 LT16 2.101.66 LT17 1.32 2.68 1.69 LT19 4.05 1.67 1.91 3.99 1.68 1.16 CT2 3.56 CT81.01 CT10 1.81 CT14 1.82 CT1 1.24 1.34 CT5 2.96 1.33 2.99 2.39 CT6 1.10CT7 1.40 CT9 1.39 1.09 CT11 2.22 1.48 2.26 1.12

[0977] Because amplification of the various DNAs described above occursin various cancerous tumors and tumor cell lines derived from varioushuman tissues, these molecules likely play a significant role in tumorformation and/or growth. As a result, amplification and/or enhancedexpression of these molecules can serve as a diagnostic for detectingthe presence of tumor in an individual and antagonists (e.g.,antibodies) directed against the proteins encoded by the above describedDNA molecules would be expected to have utility in cancer therapy.

Example 69 Gene Expression in Bovine Pericytes (Assay 105)

[0978] This assay is designed to identify gene expression patterns inpericytes induced by the hits in assay 93 described above. Bovinepericytes are plated on 60 mm culture dishes in growth media for 1 week.On day 1, various PRO polypeptides are diluted (1%) and incubated withthe pericytes for 1, 4 and 24 hr. timepoints. The cells are harvestedand the RNA isolated using TRI-Reagent following the includedinstructions. The RNA is then quantified by reading the 260/280 OD usinga spectrophotometer. The gene expression analysis is done by TaqManreactions using Perkin Elmer reagents and specially designed bovineprobes and primers. Expression of the following genes is analyzed:GAPDH, beta-integrin, connective tissue growth factor (CTGF), ICAM-1,monocyte chemoattractant protein-1 (MCP-1), osteopontin, transforminggrowth factor-beta (TGF-beta), TGF-beta receptor, tissue inhibitor ofmetalloproteinase (TIMP), tissue factor (TF), VEGF-α, thrombospondin,VEGF-β, angiopoeitin-2, and collagenase. Replicates are then averagedand the SD determined. The gene expression levels are then normalized toGAPDH. These are then normalized to the expression levels obtained witha protein (PIN32) which does not significantly induce gene expression inbovine pericytes when compared to untreated controls. Any PROpolypeptide that gives a gene expression level 2-fold or higher over thePIN32 control is considered a positive hit.

[0979] The following PRO polypeptides tested positive in this assay:PRO217.

Example 70 Cytokine Release Assay (Assay 120)

[0980] This assay is designed to determine whether PRO polypeptides ofthe present invention are capable of inducing the release of cytokinesfrom peripheral blood mononuclear cells (PBMCs). PRO polypeptidescapable of inducing the release of cytokines from PBMCs are useful fromthe treatment of conditions which would benefit from enhanced cytokinerelease and will be readily evident to those of ordinary skill in theart. Specifically, 1×10⁶ cells/ml of peripheral blood mononuclear cells(PBMC) are cultured with 1% of a PRO polypeptide for 3 days in completeRPMI media. The supernatant is then harvested and tested for increasedconcentrations of various cytokines by ELISA as compared to a human IgGtreated control. A positive in the assay is a 10-fold or greaterincrease in cytokine concentration in the PRO polypeptide treated sampleas compared to the human IgG treated control.

[0981] The following polypeptides tested positive in this assay:PRO9940.

Example 71 Identification of PRO Polypeptides That Activate Pericytes(Assay 125)

[0982] This assay shows that certain polypeptides of the invention actto activate proliferation of pericyte cells and, therefore, are usefulnot only as diagnostic markers for particular types ofpericyte-associated tumors but also for giving rise to antagonists whichwould be expected to be useful for the therapeutic treatment ofpericyte-associated tumors. Activation of pericyte proliferation alsocorrelates with the induction of angiogenesis and, as such, PROpolypeptides capable of inducing pericyte proliferation would beexpected to be useful for the treatment of conditions where inducedangiogenesis would be beneficial including, for example, wound healing,and the like. Specifically, on day 1, pericytes are received from VECTechnologies, and all but 5 ml media is removed from the flask. On day2, the pericytes are trypsinized, washed, spun and plated on 96 wellplates. On day 7, the media is removed and the pericytes are treatedwith 100 μl of either the specific PRO polypeptide or control treatments(positive control=DME+5% +/− PDGF @ 500 ng/μl; negative control=PIN32, apolypeptide determined to have no significant effect on pericyteproliferation). C-fos and GAPDH gene expression levels are thendetermined and the replicates are averaged and the SD is determined. Thec-fos values are normalized to GAPDH and the results are expressed asfold increase over PIN2. Anything providing at least a 2-fold or higherresponse as compared to the negative control is considered positive forthe assay.

[0983] The following polypeptides tested positive in this assay: PRO217.

Example 72 Identification of Receptor/Ligand Interactions

[0984] In this assay, various PRO polypeptides are tested for ability tobind to a panel of potential receptor or ligand molecules for thepurpose of identifying receptor/ligand interactions. The identificationof a ligand for a known receptor, a receptor for a known ligand or anovel receptor/ligand pair is useful for a variety of indicationsincluding, for example, targeting bioactive molecules (linked to theligand or receptor) to a cell known to express the receptor or ligand,use of the receptor or ligand as a reagent to detect the presence of theligand or receptor in a composition suspected of containing the same,wherein the composition may comprise cells suspected of expressing theligand or receptor, modulating the growth of or another biological orimmunological activity of a cell known to express or respond to thereceptor or ligand, modulating the immune response of cells or towardcells that express the receptor or ligand, allowing the preparaion ofagonists, antagonists and/or antibodies directed against the receptor orligand which will modulate the growth of or a biological orimmunological activity of a cell expressing the receptor or ligand, andvarious other indications which will be readily apparent to theordinarily skilled artisan.

[0985] The assay is performed as follows. A PRO polypeptide of thepresent invention suspected of being a ligand for a receptor isexpressed as a fusion protein containing the Fc domain of human IgG (aninmunoadhesin). Receptor-ligand binding is detected by allowinginteraction of the immunoadhesin polypeptide with cells (e.g. Cos cells)expressing candidate PRO polypeptide receptors and visualization ofbound innunoadhesin with fluorescent reagents directed toward the Fcfusion domain and examination by microscope. Cells expressing candidatereceptors are produced by transient transfection, in parallel, ofdefined subsets of a library of cDNA expression vectors encoding PROpolypeptides that may function as receptor molecules. Cells are thenincubated for 1 hour in the presence of the PRO polypeptideimmunoadhesin being tested for possible receptor binding. The cells arethen washed and fixed with paraformaldehyde. The cells are thenincubated with fluorescent conjugated antibody directed against the Fcportion of the PRO polypeptide immunoadhesin (e.g. FITC conjugated goatanti-human-Fc antibody). The cells are then washed again and examined bymicroscope. A positive interaction is judged by the presence offluorescent labeling of cells transfected with cDNA encoding aparticular PRO polypeptide receptor or pool of receptors and an absenceof similar fluorescent labeling of similarly prepared cells that havebeen transfected with other cDNA or pools of cDNA. If a defined pool ofcDNA expression vectors is judged to be positive for interaction with aPRO polypeptide inmunoadhesin, the individual cDNA species that comprisethe pool are tested individually (the pool is “broken down”) todetermine the specific cDNA that encodes a receptor able to interactwith the PRO polypeptide immunoadhesin.

[0986] In another embodiment of this assay, an epitope-tagged potentialligand PRO polypeptide (e.g. 8 histidine “His” tag) is allowed tointeract with a panel of potential receptor PRO polypeptide moleculesthat have been expressed as fusions with the Fc domain of human IgG(immunoadhesins). Following a 1 hour co-incubation with the epitopetagged PRO polypeptide, the candidate receptors are eachimmnunoprecipitated with protein A beads and the beads are washed.Potential ligand interaction is determined by western blot analysis ofthe immnunoprecipitated complexes with antibody directed towards theepitope tag. An interaction is judged to occur if a band of theanticipated molecular weight of the epitope tagged protein is observedin the western blot analysis with a candidate receptor, but is notobserved to occur with the other members of the panel of potentialreceptors.

[0987] Using these assays, the following receptor/ligand interactionshave been herein identified:

[0988] (1) PRO533 binds to the fibroblast growth factor receptor-4(FGFR-4; see Partanen et al., EMBO J. 10(6):1347-1354 (1991)).

[0989] (2) PRO301 binds to itself and, therefore, functions as anadhesion molecule.

[0990] (3) PRO187 binds to the fibroblast growth factor receptor-3(FGFR-3; see Keegan et al., Proc. Natl. Acad. Sci. USA 88:1095-1099(1991)) with high affinity and with lower affinity to to FGFR-1, 2 and 4(see Isacchi et al., Nuc. Acids Res. 18(7):1906 (1990), Dionne et al.,EMBO J. 9(9):2685-2692 (1990) and Partanen et al., EMBO J.10(6):1347-1354 (1991), respectively).

[0991] (4) PRO337 binds to PRO6004.

[0992] (5) PRO1411 binds to PRO4356.

[0993] (6) PRO10096 binds to PRO2630.

[0994] (7) PRO246 binds to itself and, therefore, functions as anadhesion molecule.

[0995] (8) PRO6307 binds to PRO265.

[0996] (9) PRO6003 binds to PRO941.

[0997] Deposit of Material

[0998] The following materials have been deposited with the AmericanType Culture Collection, 10801 University Blvd., Manassas, Va.20110-2209, USA (ATCC): TABLE 10 Material ATCC Dep. No. Deposit DateDNA22779-1130 209280 Sep. 18, 1997 DNA26846-1397 203406 Oct. 27, 1998DNA32279-1131 209259 Sep. 16, 1997 DNA32288-1132 209261 Sep. 16, 1997DNA33094-1131 209256 Sep. 16, 1997 DNA33785-1143 209417 Oct. 28, 1997DNA35663-1129 209201 Jun. 18, 1997 DNA46777-1253 209619 Feb. 5, 1998DNA60783-1611 203130 Aug. 18, 1998 DNA62306-1570 203254 Sep. 9, 1998DNA62880-1513 203097 Aug. 4, 1998 DNA64896-1539 203238 Sep. 9, 1998DNA71290-1630 203275 Sep. 22, 1998 DNA96031-2664 PTA-237 Jun. 15, 1999DNA108722-2743 PTA-552 Aug. 17, 1999 DNA35674-1142 209416 Oct. 28, 1997DNA41234 209618 Feb. 5, 1998 DNA77503-1686 203362 Oct. 20, 1998DNA49435-1219 209480 Nov. 21, 1997 DNA40628-1216 209432 Nov. 7, 1997DNA27864-1155 209375 Oct. 16, 1997 DNA43316-1237 209487 Nov. 21, 1997DNA59212-1627 203245 Sep. 9, 1998 DNA86576-2595 203868 Mar. 23, 1999DNA35639-1172 209396 Oct. 17, 1997 DNA36350-1158 209378 Oct. 16, 1997DNA53906-1368 209747 Apr. 7, 1998 DNA125185-2806 PTA-1031 Dec. 7, 1999DNA83568-2692 PTA-386 Jul. 20, 1999

[0999] These deposits were made under the provisions of the BudapestTreaty on the International Recognition of the Deposit of Microorganismsfor the Purpose of Pat. Procedure and the Regulations thereunder(Budapest Treaty). This assures maintenance of a viable culture of thedeposit for 30 years from the date of deposit. The deposits will be madeavailable by ATCC under the terms of the Budapest Treaty, and subject toan agreement between Genentech, Inc. and ATCC, which assures permanentand unrestricted availability of the progeny of the culture of thedeposit to the public upon issuance of the pertinent U.S. patent or uponlaying open to the public of any U.S. or foreign patent application,whichever comes first, and assures availability of the progeny to onedetermined by the U.S. Commissioner of Pat.s and Trademarks to beentitled thereto according to 35 USC § 122 and the Commissioner's rulespursuant thereto (including 37 CFR § 1.14 with particular reference to886 OG 638).

[1000] The assignee of the present application has agreed that if aculture of the materials on deposit should die or be lost or destroyedwhen cultivated under suitable conditions, the materials will bepromptly replaced on notification with another of the same. Availabilityof the deposited material is not to be construed as a license topractice the invention in contravention of the rights granted under theauthority of any government in accordance with its patent laws.

[1001] The foregoing written specification is considered to besufficient to enable one skilled in the art to practice the invention.The present invention is not to be limited in scope by the constructdeposited, since the deposited embodiment is intended as a singleillustration of certain aspects of the invention and any constructs thatare functionally equivalent are within the scope of this invention. Thedeposit of material herein does not constitute an admission that thewritten description herein contained is inadequate to enable thepractice of any aspect of the invention, including the best modethereof, nor is it to be construed as limiting the scope of the claimsto the specific illustrations that it represents. Indeed, variousmodifications of the invention in addition to those shown and describedherein will become apparent to those skilled in the art from theforegoing description and fall within the scope of the appended claims.

1 151 1 43 DNA Artificial Sequence Synthetic oligonucleotide probe 1tgtaaaacga cggccagtta aatagacctg caattattaa tct 43 2 41 DNA ArtificialSequence Synthetic oligonucleotide probe 2 caggaaacag ctatgaccacctgcacacct gcaaatccat t 41 3 2290 DNA Homo Sapien 3 ggctgaggggaggcccggag cctttctggg gcctggggga tcctcttgca 50 ctggtgggtg gagagaagcgcctgcagcca accagggtca ggctgtgctc 100 acagtttcct ctggcggcat gtaaaggctccacaaaggag ttgggagttc 150 aaatgaggct gctgcggacg gcctgaggat ggaccccaagccctggacct 200 gccgagcgtg gcactgaggc agcggctgac gctactgtga gggaaagaag250 gttgtgagca gccccgcagg acccctggcc agccctggcc ccagcctctg 300ccggagccct ctgtggaggc agagccagtg gagcccagtg aggcagggct 350 gcttggcagccaccggcctg caactcagga acccctccag aggccatgga 400 caggctgccc cgctgacggccagggtgaag catgtgagga gccgccccgg 450 agccaagcag gagggaagag gctttcatagattctattca caaagaataa 500 ccaccatttt gcaaggacca tgaggccact gtgcgtgacatgctggtggc 550 tcggactgct ggctgccatg ggagctgttg caggccagga ggacggtttt600 gagggcactg aggagggctc gccaagagag ttcatttacc taaacaggta 650caagcgggcg ggcgagtccc aggacaagtg cacctacacc ttcattgtgc 700 cccagcagcgggtcacgggt gccatctgcg tcaactccaa ggagcctgag 750 gtgcttctgg agaaccgagtgcataagcag gagctagagc tgctcaacaa 800 tgagctgctc aagcagaagc ggcagatcgagacgctgcag cagctggtgg 850 aggtggacgg cggcattgtg agcgaggtga agctgctgcgcaaggagagc 900 cgcaacatga actcgcgggt cacgcagctc tacatgcagc tcctgcacga950 gatcatccgc aagcgggaca acgcgttgga gctctcccag ctggagaaca 1000ggatcctgaa ccagacagcc gacatgctgc agctggccag caagtacaag 1050 gacctggagcacaagtacca gcacctggcc acactggccc acaaccaatc 1100 agagatcatc gcgcagcttgaggagcactg ccagagggtg ccctcggcca 1150 ggcccgtccc ccagccaccc cccgctgccccgccccgggt ctaccaacca 1200 cccacctaca accgcatcat caaccagatc tctaccaacgagatccagag 1250 tgaccagaac ctgaaggtgc tgccaccccc tctgcccact atgcccactc1300 tcaccagcct cccatcttcc accgacaagc cgtcgggccc atggagagac 1350tgcctgcagg ccctggagga tggccacgac accagctcca tctacctggt 1400 gaagccggagaacaccaacc gcctcatgca ggtgtggtgc gaccagagac 1450 acgaccccgg gggctggaccgtcatccaga gacgcctgga tggctctgtt 1500 aacttcttca ggaactggga gacgtacaagcaagggtttg ggaacattga 1550 cggcgaatac tggctgggcc tggagaacat ttactggctgacgaaccaag 1600 gcaactacaa actcctggtg accatggagg actggtccgg ccgcaaagtc1650 tttgcagaat acgccagttt ccgcctggaa cctgagagcg agtattataa 1700gctgcggctg gggcgctacc atggcaatgc gggtgactcc tttacatggc 1750 acaacggcaagcagttcacc accctggaca gagatcatga tgtctacaca 1800 ggaaactgtg cccactaccagaagggaggc tggtggtata acgcctgtgc 1850 ccactccaac ctcaacgggg tctggtaccgcgggggccat taccggagcc 1900 gctaccagga cggagtctac tgggctgagt tccgaggaggctcttactca 1950 ctcaagaaag tggtgatgat gatccgaccg aaccccaaca ccttccacta2000 agccagctcc ccctcctgac ctctcgtggc cattgccagg agcccaccct 2050ggtcacgctg gccacagcac aaagaacaac tcctcaccag ttcatcctga 2100 ggctgggaggaccgggatgc tggattctgt tttccgaagt cactgcagcg 2150 gatgatggaa ctgaatcgatacggtgtttt ctgtccctcc tactttcctt 2200 cacaccagac agcccctcat gtctccaggacaggacagga ctacagacaa 2250 ctctttcttt aaataaatta agtctctaca ataaaaaaaa2290 4 493 PRT Homo Sapien 4 Met Arg Pro Leu Cys Val Thr Cys Trp Trp LeuGly Leu Leu Ala 1 5 10 15 Ala Met Gly Ala Val Ala Gly Gln Glu Asp GlyPhe Glu Gly Thr 20 25 30 Glu Glu Gly Ser Pro Arg Glu Phe Ile Tyr Leu AsnArg Tyr Lys 35 40 45 Arg Ala Gly Glu Ser Gln Asp Lys Cys Thr Tyr Thr PheIle Val 50 55 60 Pro Gln Gln Arg Val Thr Gly Ala Ile Cys Val Asn Ser LysGlu 65 70 75 Pro Glu Val Leu Leu Glu Asn Arg Val His Lys Gln Glu Leu Glu80 85 90 Leu Leu Asn Asn Glu Leu Leu Lys Gln Lys Arg Gln Ile Glu Thr 95100 105 Leu Gln Gln Leu Val Glu Val Asp Gly Gly Ile Val Ser Glu Val 110115 120 Lys Leu Leu Arg Lys Glu Ser Arg Asn Met Asn Ser Arg Val Thr 125130 135 Gln Leu Tyr Met Gln Leu Leu His Glu Ile Ile Arg Lys Arg Asp 140145 150 Asn Ala Leu Glu Leu Ser Gln Leu Glu Asn Arg Ile Leu Asn Gln 155160 165 Thr Ala Asp Met Leu Gln Leu Ala Ser Lys Tyr Lys Asp Leu Glu 170175 180 His Lys Tyr Gln His Leu Ala Thr Leu Ala His Asn Gln Ser Glu 185190 195 Ile Ile Ala Gln Leu Glu Glu His Cys Gln Arg Val Pro Ser Ala 200205 210 Arg Pro Val Pro Gln Pro Pro Pro Ala Ala Pro Pro Arg Val Tyr 215220 225 Gln Pro Pro Thr Tyr Asn Arg Ile Ile Asn Gln Ile Ser Thr Asn 230235 240 Glu Ile Gln Ser Asp Gln Asn Leu Lys Val Leu Pro Pro Pro Leu 245250 255 Pro Thr Met Pro Thr Leu Thr Ser Leu Pro Ser Ser Thr Asp Lys 260265 270 Pro Ser Gly Pro Trp Arg Asp Cys Leu Gln Ala Leu Glu Asp Gly 275280 285 His Asp Thr Ser Ser Ile Tyr Leu Val Lys Pro Glu Asn Thr Asn 290295 300 Arg Leu Met Gln Val Trp Cys Asp Gln Arg His Asp Pro Gly Gly 305310 315 Trp Thr Val Ile Gln Arg Arg Leu Asp Gly Ser Val Asn Phe Phe 320325 330 Arg Asn Trp Glu Thr Tyr Lys Gln Gly Phe Gly Asn Ile Asp Gly 335340 345 Glu Tyr Trp Leu Gly Leu Glu Asn Ile Tyr Trp Leu Thr Asn Gln 350355 360 Gly Asn Tyr Lys Leu Leu Val Thr Met Glu Asp Trp Ser Gly Arg 365370 375 Lys Val Phe Ala Glu Tyr Ala Ser Phe Arg Leu Glu Pro Glu Ser 380385 390 Glu Tyr Tyr Lys Leu Arg Leu Gly Arg Tyr His Gly Asn Ala Gly 395400 405 Asp Ser Phe Thr Trp His Asn Gly Lys Gln Phe Thr Thr Leu Asp 410415 420 Arg Asp His Asp Val Tyr Thr Gly Asn Cys Ala His Tyr Gln Lys 425430 435 Gly Gly Trp Trp Tyr Asn Ala Cys Ala His Ser Asn Leu Asn Gly 440445 450 Val Trp Tyr Arg Gly Gly His Tyr Arg Ser Arg Tyr Gln Asp Gly 455460 465 Val Tyr Trp Ala Glu Phe Arg Gly Gly Ser Tyr Ser Leu Lys Lys 470475 480 Val Val Met Met Ile Arg Pro Asn Pro Asn Thr Phe His 485 490 5 33DNA Artificial Sequence Synthetic oligonucleotide probe 5 gctgacgaaccaaggcaact acaaactcct ggt 33 6 41 DNA Artificial Sequence Syntheticoligonucleotide probe 6 tgcggccgga ccagtcctcc atggtcacca ggagtttgta g 417 33 DNA Artificial Sequence Synthetic oligonucleotide probe 7ggtggtgaac tgcttgccgt tgtgccatgt aaa 33 8 1218 DNA Homo Sapien 8cccacgcgtc cggcgccgtg gcctcgcgtc catctttgcc gttctctcgg 50 acctgtcacaaaggagtcgc gccgccgccg ccgccccctc cctccggtgg 100 gcccgggagg tagagaaagtcagtgccaca gcccgaccgc gctgctctga 150 gccctgggca cgcggaacgg gagggagtctgagggttggg gacgtctgtg 200 agggagggga acagccgctc gagcctgggg cgggcggaccggactggggc 250 cggggtaggc tctggaaagg gcccgggaga gaggtggcgt tggtcagaac300 ctgagaaaca gccgagaggt tttccaccga ggcccgcgct tgagggatct 350gaagaggttc ctagaagagg gtgttccctc tttcgggggt cctcaccaga 400 agaggttcttgggggtcgcc cttctgagga ggctgcggct aacagggccc 450 agaactgcca ttggatgtccagaatcccct gtagttgata atgttgggaa 500 taagctctgc aactttcttt ggcattcagttgttaaaaac aaataggatg 550 caaattcctc aactccaggt tatgaaaaca gtacttggaaaactgaaaac 600 tacctaaatg atcgtctttg gttgggccgt gttcttagcg agcagaagcc650 ttggccaggg tctgttgttg actctcgaag agcacatagc ccacttccta 700gggactggag gtgccgctac taccatgggt aattcctgta tctgccgaga 750 tgacagtggaacagatgaca gtgttgacac ccaacagcaa caggccgaga 800 acagtgcagt acccactgctgacacaagga gccaaccacg ggaccctgtt 850 cggccaccaa ggaggggccg aggacctcatgagccaagga gaaagaaaca 900 aaatgtggat gggctagtgt tggacacact ggcagtaatacggactcttg 950 tagataagta agtatctgac tcacggtcac ctccagtgga atgaaaagtg1000 ttctgcccgg aaccatgact ttaggactcc ttcagttcct ttaggacata 1050ctcgccaagc cttgtgctca cagggcaaag gagaatattt taatgctccg 1100 ctgatggcagagtaaatgat aagatttgat gtttttgctt gctgtcatct 1150 actttgtctg gaaatgtctaaatgtttctg tagcagaaaa cacgataaag 1200 ctatgatctt tattagag 1218 9 117 PRTHomo Sapien 9 Met Ile Val Phe Gly Trp Ala Val Phe Leu Ala Ser Arg SerLeu 1 5 10 15 Gly Gln Gly Leu Leu Leu Thr Leu Glu Glu His Ile Ala HisPhe 20 25 30 Leu Gly Thr Gly Gly Ala Ala Thr Thr Met Gly Asn Ser Cys Ile35 40 45 Cys Arg Asp Asp Ser Gly Thr Asp Asp Ser Val Asp Thr Gln Gln 5055 60 Gln Gln Ala Glu Asn Ser Ala Val Pro Thr Ala Asp Thr Arg Ser 65 7075 Gln Pro Arg Asp Pro Val Arg Pro Pro Arg Arg Gly Arg Gly Pro 80 85 90His Glu Pro Arg Arg Lys Lys Gln Asn Val Asp Gly Leu Val Leu 95 100 105Asp Thr Leu Ala Val Ile Arg Thr Leu Val Asp Lys 110 115 10 1231 DNA HomoSapien 10 cccacgcgtc cgcgcagtcg cgcagttctg cctccgcctg ccagtctcgc 50ccgcgatccc ggcccggggc tgtggcgtcg actccgaccc aggcagccag 100 cagcccgcgcgggagccgga ccgccgccgg aggagctcgg acggcatgct 150 gagccccctc ctttgctgaagcccgagtgc ggagaagccc gggcaaacgc 200 aggctaagga gaccaaagcg gcgaagtcgcgagacagcgg acaagcagcg 250 gaggagaagg aggaggaggc gaacccagag aggggcagcaaaagaagcgg 300 tggtggtggg cgtcgtggcc atggcggcgg ctatcgccag ctcgctcatc350 cgtcagaaga ggcaagcccg cgagcgcgag aaatccaacg cctgcaagtg 400tgtcagcagc cccagcaaag gcaagaccag ctgcgacaaa aacaagttaa 450 atgtcttttcccgggtcaaa ctcttcggct ccaagaagag gcgcagaaga 500 agaccagagc ctcagcttaagggtatagtt accaagctat acagccgaca 550 aggctaccac ttgcagctgc aggcggatggaaccattgat ggcaccaaag 600 atgaggacag cacttacact ctgtttaacc tcatccctgtgggtctgcga 650 gtggtggcta tccaaggagt tcaaaccaag ctgtacttgg caatgaacag700 tgagggatac ttgtacacct cggaactttt cacacctgag tgcaaattca 750aagaatcagt gtttgaaaat tattatgtga catattcatc aatgatatac 800 cgtcagcagcagtcaggccg agggtggtat ctgggtctga acaaagaagg 850 agagatcatg aaaggcaaccatgtgaagaa gaacaagcct gcagctcatt 900 ttctgcctaa accactgaaa gtggccatgtacaaggagcc atcactgcac 950 gatctcacgg agttctcccg atctggaagc gggaccccaaccaagagcag 1000 aagtgtctct ggcgtgctga acggaggcaa atccatgagc cacaatgaat1050 caacgtagcc agtgagggca aaagaagggc tctgtaacag aaccttacct 1100ccaggtgctg ttgaattctt ctagcagtcc ttcacccaaa agttcaaatt 1150 tgtcagtgacatttaccaaa caaacaggca gagttcacta ttctatctgc 1200 cattagacct tcttatcatccatactaaag c 1231 11 245 PRT Homo Sapien 11 Met Ala Ala Ala Ile Ala SerSer Leu Ile Arg Gln Lys Arg Gln 1 5 10 15 Ala Arg Glu Arg Glu Lys SerAsn Ala Cys Lys Cys Val Ser Ser 20 25 30 Pro Ser Lys Gly Lys Thr Ser CysAsp Lys Asn Lys Leu Asn Val 35 40 45 Phe Ser Arg Val Lys Leu Phe Gly SerLys Lys Arg Arg Arg Arg 50 55 60 Arg Pro Glu Pro Gln Leu Lys Gly Ile ValThr Lys Leu Tyr Ser 65 70 75 Arg Gln Gly Tyr His Leu Gln Leu Gln Ala AspGly Thr Ile Asp 80 85 90 Gly Thr Lys Asp Glu Asp Ser Thr Tyr Thr Leu PheAsn Leu Ile 95 100 105 Pro Val Gly Leu Arg Val Val Ala Ile Gln Gly ValGln Thr Lys 110 115 120 Leu Tyr Leu Ala Met Asn Ser Glu Gly Tyr Leu TyrThr Ser Glu 125 130 135 Leu Phe Thr Pro Glu Cys Lys Phe Lys Glu Ser ValPhe Glu Asn 140 145 150 Tyr Tyr Val Thr Tyr Ser Ser Met Ile Tyr Arg GlnGln Gln Ser 155 160 165 Gly Arg Gly Trp Tyr Leu Gly Leu Asn Lys Glu GlyGlu Ile Met 170 175 180 Lys Gly Asn His Val Lys Lys Asn Lys Pro Ala AlaHis Phe Leu 185 190 195 Pro Lys Pro Leu Lys Val Ala Met Tyr Lys Glu ProSer Leu His 200 205 210 Asp Leu Thr Glu Phe Ser Arg Ser Gly Ser Gly ThrPro Thr Lys 215 220 225 Ser Arg Ser Val Ser Gly Val Leu Asn Gly Gly LysSer Met Ser 230 235 240 His Asn Glu Ser Thr 245 12 744 DNA Homo Sapien12 atggccgcgg ccatcgctag cggcttgatc cgccagaagc ggcaggcgcg 50 ggagcagcactgggaccggc cgtctgccag caggaggcgg agcagcccca 100 gcaagaaccg cgggctctgcaacggcaacc tggtggatat cttctccaaa 150 gtgcgcatct tcggcctcaa gaagcgcaggttgcggcgcc aagatcccca 200 gctcaagggt atagtgacca ggttatattg caggcaaggctactacttgc 250 aaatgcaccc cgatggagct ctcgatggaa ccaaggatga cagcactaat300 tctacactct tcaacctcat accagtggga ctacgtgttg ttgccatcca 350gggagtgaaa acagggttgt atatagccat gaatggagaa ggttacctct 400 acccatcagaactttttacc cctgaatgca agtttaaaga atctgttttt 450 gaaaattatt atgtaatctactcatccatg ttgtacagac aacaggaatc 500 tggtagagcc tggtttttgg gattaaataaggaagggcaa gctatgaaag 550 ggaacagagt aaagaaaacc aaaccagcag ctcattttctacccaagcca 600 ttggaagttg ccatgtaccg agaaccatct ttgcatgatg ttggggaaac650 ggtcccgaag cctggggtga cgccaagtaa aagcacaagt gcgtctgcaa 700taatgaatgg aggcaaacca gtcaacaaga gtaagacaac atag 744 13 247 PRT HomoSapien 13 Met Ala Ala Ala Ile Ala Ser Gly Leu Ile Arg Gln Lys Arg Gln 15 10 15 Ala Arg Glu Gln His Trp Asp Arg Pro Ser Ala Ser Arg Arg Arg 2025 30 Ser Ser Pro Ser Lys Asn Arg Gly Leu Cys Asn Gly Asn Leu Val 35 4045 Asp Ile Phe Ser Lys Val Arg Ile Phe Gly Leu Lys Lys Arg Arg 50 55 60Leu Arg Arg Gln Asp Pro Gln Leu Lys Gly Ile Val Thr Arg Leu 65 70 75 TyrCys Arg Gln Gly Tyr Tyr Leu Gln Met His Pro Asp Gly Ala 80 85 90 Leu AspGly Thr Lys Asp Asp Ser Thr Asn Ser Thr Leu Phe Asn 95 100 105 Leu IlePro Val Gly Leu Arg Val Val Ala Ile Gln Gly Val Lys 110 115 120 Thr GlyLeu Tyr Ile Ala Met Asn Gly Glu Gly Tyr Leu Tyr Pro 125 130 135 Ser GluLeu Phe Thr Pro Glu Cys Lys Phe Lys Glu Ser Val Phe 140 145 150 Glu AsnTyr Tyr Val Ile Tyr Ser Ser Met Leu Tyr Arg Gln Gln 155 160 165 Glu SerGly Arg Ala Trp Phe Leu Gly Leu Asn Lys Glu Gly Gln 170 175 180 Ala MetLys Gly Asn Arg Val Lys Lys Thr Lys Pro Ala Ala His 185 190 195 Phe LeuPro Lys Pro Leu Glu Val Ala Met Tyr Arg Glu Pro Ser 200 205 210 Leu HisAsp Val Gly Glu Thr Val Pro Lys Pro Gly Val Thr Pro 215 220 225 Ser LysSer Thr Ser Ala Ser Ala Ile Met Asn Gly Gly Lys Pro 230 235 240 Val AsnLys Ser Lys Thr Thr 245 14 2609 DNA Homo Sapien 14 ctcgcagccg agcgcggccggggaagggct ctccttccag cgccgagcac 50 tgggccctgg cagacgcccc aagattgttgtgaggagtct agccagttgg 100 tgagcgctgt aatctgaacc agctgtgtcc agactgaggccccatttgca 150 ttgtttaaca tacttagaaa atgaagtgtt catttttaac attcctcctc200 caattggttt aatgctgaat tactgaagag ggctaagcaa aaccaggtgc 250ttgcgctgag ggctctgcag tggctgggag gaccccggcg ctctccccgt 300 gtcctctccacgactcgctc ggcccctctg gaataaaaca cccgcgagcc 350 ccgagggccc agaggaggccgacgtgcccg agctcctccg ggggtcccgc 400 ccgcgagctt tcttctcgcc ttcgcatctcctcctcgcgc gtcttggaca 450 tgccaggaat aaaaaggata ctcactgtta ccattctggctctctgtctt 500 ccaagccctg ggaatgcaca ggcacagtgc acgaatggct ttgacctgga550 tcgccagtca ggacagtgtt tagatattga tgaatgccga accatccccg 600aggcctgccg aggagacatg atgtgtgtta accaaaatgg cgggtattta 650 tgcattccccggacaaaccc tgtgtatcga gggccctact cgaaccccta 700 ctcgaccccc tactcaggtccgtacccagc agctgcccca ccactctcag 750 ctccaaacta tcccacgatc tccaggcctcttatatgccg ctttggatac 800 cagatggatg aaagcaacca atgtgtggat gtggacgagtgtgcaacaga 850 ttcccaccag tgcaacccca cccagatctg catcaatact gaaggcgggt900 acacctgctc ctgcaccgac ggatattggc ttctggaagg ccagtgctta 950gacattgatg aatgtcgcta tggttactgc cagcagctct gtgcgaatgt 1000 tcctggatcctattcttgta catgcaaccc tggttttacc ctcaatgagg 1050 atggaaggtc ttgccaagatgtgaacgagt gtgccaccga gaacccctgc 1100 gtgcaaacct gcgtcaacac ctacggctctctcatctgcc gctgtgaccc 1150 aggatatgaa cttgaggaag atggcgttca ttgcagtgatatggacgagt 1200 gcagcttctc tgagttcctc tgccaacatg agtgtgtgaa ccagcccggc1250 acatacttct gctcctgccc tccaggctac atcctgctgg atgacaaccg 1300aagctgccaa gacatcaacg aatgtgagca caggaaccac acgtgcaacc 1350 tgcagcagacgtgctacaat ttacaagggg gcttcaaatg catcgacccc 1400 atccgctgtg aggagccttatctgaggatc agtgataacc gctgtatgtg 1450 tcctgctgag aaccctggct gcagagaccagccctttacc atcttgtacc 1500 gggacatgga cgtggtgtca ggacgctccg ttcccgctgacatcttccaa 1550 atgcaagcca cgacccgcta ccctggggcc tattacattt tccagatcaa1600 atctgggaat gagggcagag aattttacat gcggcaaacg ggccccatca 1650gtgccaccct ggtgatgaca cgccccatca aagggccccg ggaaatccag 1700 ctggacttggaaatgatcac tgtcaacact gtcatcaact tcagaggcag 1750 ctccgtgatc cgactgcggatatatgtgtc gcagtaccca ttctgagcct 1800 cgggctggag cctccgacgc tgcctctcattggcaccaag ggacaggaga 1850 agagaggaaa taacagagag aatgagagcg acacagacgttaggcatttc 1900 ctgctgaacg tttccccgaa gagtcagccc cgacttcctg actctcacct1950 gtactattgc agacctgtca ccctgcagga cttgccaccc ccagttccta 2000tgacacagtt atcaaaaagt attatcattg ctcccctgat agaagattgt 2050 tggtgaattttcaaggcctt cagtttattt ccactatttt caaagaaaat 2100 agattaggtt tgcgggggtctgagtctatg ttcaaagact gtgaacagct 2150 tgctgtcact tcttcacctc ttccactccttctctcactg tgttactgct 2200 ttgcaaagac ccgggagctg gcggggaacc ctgggagtagctagtttgct 2250 ttttgcgtac acagagaagg ctatgtaaac aaaccacagc aggatcgaag2300 ggtttttaga gaatgtgttt caaaaccatg cctggtattt tcaaccataa 2350aagaagtttc agttgtcctt aaatttgtat aacggtttaa ttctgtcttg 2400 ttcattttgagtatttttaa aaaatatgtc gtagaattcc ttcgaaaggc 2450 cttcagacac atgctatgttctgtcttccc aaacccagtc tcctctccat 2500 tttagcccag tgttttcttt gaggaccccttaatcttgct ttctttagaa 2550 tttttaccca attggattgg aatgcagagg tctccaaactgattaaatat 2600 ttgaagaga 2609 15 448 PRT Homo Sapien 15 Met Pro Gly IleLys Arg Ile Leu Thr Val Thr Ile Leu Ala Leu 1 5 10 15 Cys Leu Pro SerPro Gly Asn Ala Gln Ala Gln Cys Thr Asn Gly 20 25 30 Phe Asp Leu Asp ArgGln Ser Gly Gln Cys Leu Asp Ile Asp Glu 35 40 45 Cys Arg Thr Ile Pro GluAla Cys Arg Gly Asp Met Met Cys Val 50 55 60 Asn Gln Asn Gly Gly Tyr LeuCys Ile Pro Arg Thr Asn Pro Val 65 70 75 Tyr Arg Gly Pro Tyr Ser Asn ProTyr Ser Thr Pro Tyr Ser Gly 80 85 90 Pro Tyr Pro Ala Ala Ala Pro Pro LeuSer Ala Pro Asn Tyr Pro 95 100 105 Thr Ile Ser Arg Pro Leu Ile Cys ArgPhe Gly Tyr Gln Met Asp 110 115 120 Glu Ser Asn Gln Cys Val Asp Val AspGlu Cys Ala Thr Asp Ser 125 130 135 His Gln Cys Asn Pro Thr Gln Ile CysIle Asn Thr Glu Gly Gly 140 145 150 Tyr Thr Cys Ser Cys Thr Asp Gly TyrTrp Leu Leu Glu Gly Gln 155 160 165 Cys Leu Asp Ile Asp Glu Cys Arg TyrGly Tyr Cys Gln Gln Leu 170 175 180 Cys Ala Asn Val Pro Gly Ser Tyr SerCys Thr Cys Asn Pro Gly 185 190 195 Phe Thr Leu Asn Glu Asp Gly Arg SerCys Gln Asp Val Asn Glu 200 205 210 Cys Ala Thr Glu Asn Pro Cys Val GlnThr Cys Val Asn Thr Tyr 215 220 225 Gly Ser Leu Ile Cys Arg Cys Asp ProGly Tyr Glu Leu Glu Glu 230 235 240 Asp Gly Val His Cys Ser Asp Met AspGlu Cys Ser Phe Ser Glu 245 250 255 Phe Leu Cys Gln His Glu Cys Val AsnGln Pro Gly Thr Tyr Phe 260 265 270 Cys Ser Cys Pro Pro Gly Tyr Ile LeuLeu Asp Asp Asn Arg Ser 275 280 285 Cys Gln Asp Ile Asn Glu Cys Glu HisArg Asn His Thr Cys Asn 290 295 300 Leu Gln Gln Thr Cys Tyr Asn Leu GlnGly Gly Phe Lys Cys Ile 305 310 315 Asp Pro Ile Arg Cys Glu Glu Pro TyrLeu Arg Ile Ser Asp Asn 320 325 330 Arg Cys Met Cys Pro Ala Glu Asn ProGly Cys Arg Asp Gln Pro 335 340 345 Phe Thr Ile Leu Tyr Arg Asp Met AspVal Val Ser Gly Arg Ser 350 355 360 Val Pro Ala Asp Ile Phe Gln Met GlnAla Thr Thr Arg Tyr Pro 365 370 375 Gly Ala Tyr Tyr Ile Phe Gln Ile LysSer Gly Asn Glu Gly Arg 380 385 390 Glu Phe Tyr Met Arg Gln Thr Gly ProIle Ser Ala Thr Leu Val 395 400 405 Met Thr Arg Pro Ile Lys Gly Pro ArgGlu Ile Gln Leu Asp Leu 410 415 420 Glu Met Ile Thr Val Asn Thr Val IleAsn Phe Arg Gly Ser Ser 425 430 435 Val Ile Arg Leu Arg Ile Tyr Val SerGln Tyr Pro Phe 440 445 16 2447 DNA Homo Sapien 16 caggtccaac tgcacctcggttctatcgat tgaattcccc ggggatcctc 50 tagagatccc tcgacctcga cccacgcgtccgaacacagg tccttgttgc 100 tgcagagaag cagttgtttt gctggaagga gggagtgcgcgggctgcccc 150 gggctcctcc ctgccgcctc ctctcagtgg atggttccag gcaccctgtc200 tggggcaggg agggcacagg cctgcacatc gaaggtgggg tgggaccagg 250ctgcccctcg ccccagcatc caagtcctcc cttgggcgcc cgtggccctg 300 cagactctcagggctaaggt cctctgttgc tttttggttc caccttagaa 350 gaggctccgc ttgactaagagtagcttgaa ggaggcacca tgcaggagct 400 gcatctgctc tggtgggcgc ttctcctgggcctggctcag gcctgccctg 450 agccctgcga ctgtggggaa aagtatggct tccagatcgccgactgtgcc 500 taccgcgacc tagaatccgt gccgcctggc ttcccggcca atgtgactac550 actgagcctg tcagccaacc ggctgccagg cttgccggag ggtgccttca 600gggaggtgcc cctgctgcag tcgctgtggc tggcacacaa tgagatccgc 650 acggtggccgccggagccct ggcctctctg agccatctca agagcctgga 700 cctcagccac aatctcatctctgactttgc ctggagcgac ctgcacaacc 750 tcagtgccct ccaattgctc aagatggacagcaacgagct gaccttcatc 800 ccccgcgacg ccttccgcag cctccgtgct ctgcgctcgctgcaactcaa 850 ccacaaccgc ttgcacacat tggccgaggg caccttcacc ccgctcaccg900 cgctgtccca cctgcagatc aacgagaacc ccttcgactg cacctgcggc 950atcgtgtggc tcaagacatg ggccctgacc acggccgtgt ccatcccgga 1000 gcaggacaacatcgcctgca cctcacccca tgtgctcaag ggtacaccgc 1050 tgagccgcct gccgccactgccatgctcgg cgccctcagt gcagctcagc 1100 taccaaccca gccaggatgg tgccgagctgcggcctggtt ttgtgctggc 1150 actgcactgt gatgtggacg ggcagccggc ccctcagcttcactggcaca 1200 tccagatacc cagtggcatt gtggagatca ccagccccaa cgtgggcact1250 gatgggcgtg ccctgcctgg cacccctgtg gccagctccc agccgcgctt 1300ccaggccttt gccaatggca gcctgcttat ccccgacttt ggcaagctgg 1350 aggaaggcacctacagctgc ctggccacca atgagctggg cagtgctgag 1400 agctcagtgg acgtggcactggccacgccc ggtgagggtg gtgaggacac 1450 actggggcgc aggttccatg gcaaagcggttgagggaaag ggctgctata 1500 cggttgacaa cgaggtgcag ccatcagggc cggaggacaatgtggtcatc 1550 atctacctca gccgtgctgg gaaccctgag gctgcagtcg cagaaggggt1600 ccctgggcag ctgcccccag gcctgctcct gctgggccaa agcctcctcc 1650tcttcttctt cctcacctcc ttctagcccc acccagggct tccctaactc 1700 ctccccttgcccctaccaat gcccctttaa gtgctgcagg ggtctggggt 1750 tggcaactcc tgaggcctgcatgggtgact tcacattttc ctacctctcc 1800 ttctaatctc ttctagagca cctgctatccccaacttcta gacctgctcc 1850 aaactagtga ctaggataga atttgatccc ctaactcactgtctgcggtg 1900 ctcattgctg ctaacagcat tgcctgtgct ctcctctcag gggcagcatg1950 ctaacggggc gacgtcctaa tccaactggg agaagcctca gtggtggaat 2000tccaggcact gtgactgtca agctggcaag ggccaggatt gggggaatgg 2050 agctggggcttagctgggag gtggtctgaa gcagacaggg aatgggagag 2100 gaggatggga agtagacagtggctggtatg gctctgaggc tccctggggc 2150 ctgctcaagc tcctcctgct ccttgctgttttctgatgat ttgggggctt 2200 gggagtccct ttgtcctcat ctgagactga aatgtggggatccaggatgg 2250 ccttccttcc tcttaccctt cctccctcag cctgcaacct ctatcctgga2300 acctgtcctc cctttctccc caactatgca tctgttgtct gctcctctgc 2350aaaggccagc cagcttggga gcagcagaga aataaacagc atttctgatg 2400 ccaaaaaaaaaaaaaaaaaa gggcggccgc gactctagag tcgacct 2447 17 428 PRT Homo Sapien 17Met Gln Glu Leu His Leu Leu Trp Trp Ala Leu Leu Leu Gly Leu 1 5 10 15Ala Gln Ala Cys Pro Glu Pro Cys Asp Cys Gly Glu Lys Tyr Gly 20 25 30 PheGln Ile Ala Asp Cys Ala Tyr Arg Asp Leu Glu Ser Val Pro 35 40 45 Pro GlyPhe Pro Ala Asn Val Thr Thr Leu Ser Leu Ser Ala Asn 50 55 60 Arg Leu ProGly Leu Pro Glu Gly Ala Phe Arg Glu Val Pro Leu 65 70 75 Leu Gln Ser LeuTrp Leu Ala His Asn Glu Ile Arg Thr Val Ala 80 85 90 Ala Gly Ala Leu AlaSer Leu Ser His Leu Lys Ser Leu Asp Leu 95 100 105 Ser His Asn Leu IleSer Asp Phe Ala Trp Ser Asp Leu His Asn 110 115 120 Leu Ser Ala Leu GlnLeu Leu Lys Met Asp Ser Asn Glu Leu Thr 125 130 135 Phe Ile Pro Arg AspAla Phe Arg Ser Leu Arg Ala Leu Arg Ser 140 145 150 Leu Gln Leu Asn HisAsn Arg Leu His Thr Leu Ala Glu Gly Thr 155 160 165 Phe Thr Pro Leu ThrAla Leu Ser His Leu Gln Ile Asn Glu Asn 170 175 180 Pro Phe Asp Cys ThrCys Gly Ile Val Trp Leu Lys Thr Trp Ala 185 190 195 Leu Thr Thr Ala ValSer Ile Pro Glu Gln Asp Asn Ile Ala Cys 200 205 210 Thr Ser Pro His ValLeu Lys Gly Thr Pro Leu Ser Arg Leu Pro 215 220 225 Pro Leu Pro Cys SerAla Pro Ser Val Gln Leu Ser Tyr Gln Pro 230 235 240 Ser Gln Asp Gly AlaGlu Leu Arg Pro Gly Phe Val Leu Ala Leu 245 250 255 His Cys Asp Val AspGly Gln Pro Ala Pro Gln Leu His Trp His 260 265 270 Ile Gln Ile Pro SerGly Ile Val Glu Ile Thr Ser Pro Asn Val 275 280 285 Gly Thr Asp Gly ArgAla Leu Pro Gly Thr Pro Val Ala Ser Ser 290 295 300 Gln Pro Arg Phe GlnAla Phe Ala Asn Gly Ser Leu Leu Ile Pro 305 310 315 Asp Phe Gly Lys LeuGlu Glu Gly Thr Tyr Ser Cys Leu Ala Thr 320 325 330 Asn Glu Leu Gly SerAla Glu Ser Ser Val Asp Val Ala Leu Ala 335 340 345 Thr Pro Gly Glu GlyGly Glu Asp Thr Leu Gly Arg Arg Phe His 350 355 360 Gly Lys Ala Val GluGly Lys Gly Cys Tyr Thr Val Asp Asn Glu 365 370 375 Val Gln Pro Ser GlyPro Glu Asp Asn Val Val Ile Ile Tyr Leu 380 385 390 Ser Arg Ala Gly AsnPro Glu Ala Ala Val Ala Glu Gly Val Pro 395 400 405 Gly Gln Leu Pro ProGly Leu Leu Leu Leu Gly Gln Ser Leu Leu 410 415 420 Leu Phe Phe Phe LeuThr Ser Phe 425 18 22 DNA Artificial Sequence Synthetic oligonucleotideprobe 18 gtggctggca cacaatgaga tc 22 19 22 DNA Artificial SequenceSynthetic oligonucleotide probe 19 ccaatgtgtg caagcggttg tg 22 20 50 DNAArtificial Sequence Synthetic oligonucleotide probe 20 tcaagagcctggacctcagc cacaatctca tctctgactt tgcctggagc 50 21 2033 DNA Homo Sapien21 ccaggccggg aggcgacgcg cccagccgtc taaacgggaa cagccctggc 50 tgagggagctgcagcgcagc agagtatctg acggcgccag gttgcgtagg 100 tgcggcacga ggagttttcccggcagcgag gaggtcctga gcagcatggc 150 ccggaggagc gccttccctg ccgccgcgctctggctctgg agcatcctcc 200 tgtgcctgct ggcactgcgg gcggaggccg ggccgccgcaggaggagagc 250 ctgtacctat ggatcgatgc tcaccaggca agagtactca taggatttga300 agaagatatc ctgattgttt cagaggggaa aatggcacct tttacacatg 350atttcagaaa agcgcaacag agaatgccag ctattcctgt caatatccat 400 tccatgaattttacctggca agctgcaggg caggcagaat acttctatga 450 attcctgtcc ttgcgctccctggataaagg catcatggca gatccaaccg 500 tcaatgtccc tctgctggga acagtgcctcacaaggcatc agttgttcaa 550 gttggtttcc catgtcttgg aaaacaggat ggggtggcagcatttgaagt 600 ggatgtgatt gttatgaatt ctgaaggcaa caccattctc caaacacctc650 aaaatgctat cttctttaaa acatgtcaac aagctgagtg cccaggcggg 700tgccgaaatg gaggcttttg taatgaaaga cgcatctgcg agtgtcctga 750 tgggttccacggacctcact gtgagaaagc cctttgtacc ccacgatgta 800 tgaatggtgg actttgtgtgactcctggtt tctgcatctg cccacctgga 850 ttctatggag tgaactgtga caaagcaaactgctcaacca cctgctttaa 900 tggagggacc tgtttctacc ctggaaaatg tatttgccctccaggactag 950 agggagagca gtgtgaaatc agcaaatgcc cacaaccctg tcgaaatgga1000 ggtaaatgca ttggtaaaag caaatgtaag tgttccaaag gttaccaggg 1050agacctctgt tcaaagcctg tctgcgagcc tggctgtggt gcacatggaa 1100 cctgccatgaacccaacaaa tgccaatgtc aagaaggttg gcatggaaga 1150 cactgcaata aaaggtacgaagccagcctc atacatgccc tgaggccagc 1200 aggcgcccag ctcaggcagc acacgccttcacttaaaaag gccgaggagc 1250 ggcgggatcc acctgaatcc aattacatct ggtgaactccgacatctgaa 1300 acgttttaag ttacaccaag ttcatagcct ttgttaacct ttcatgtgtt1350 gaatgttcaa ataatgttca ttacacttaa gaatactggc ctgaatttta 1400ttagcttcat tataaatcac tgagctgata tttactcttc cttttaagtt 1450 ttctaagtacgtctgtagca tgatggtata gattttcttg tttcagtgct 1500 ttgggacaga ttttatattatgtcaattga tcaggttaaa attttcagtg 1550 tgtagttggc agatattttc aaaattacaatgcatttatg gtgtctgggg 1600 gcaggggaac atcagaaagg ttaaattggg caaaaatgcgtaagtcacaa 1650 gaatttggat ggtgcagtta atgttgaagt tacagcattt cagattttat1700 tgtcagatat ttagatgttt gttacatttt taaaaattgc tcttaatttt 1750taaactctca atacaatata ttttgacctt accattattc cagagattca 1800 gtattaaaaaaaaaaaaatt acactgtggt agtggcattt aaacaatata 1850 atatattcta aacacaatgaaatagggaat ataatgtatg aactttttgc 1900 attggcttga agcaatataa tatattgtaaacaaaacaca gctcttacct 1950 aataaacatt ttatactgtt tgtatgtata aaataaaggtgctgctttag 2000 ttttttggaa aaaaaaaaaa aaaaaaaaaa aaa 2033 22 379 PRTHomo Sapien 22 Met Ala Arg Arg Ser Ala Phe Pro Ala Ala Ala Leu Trp LeuTrp 1 5 10 15 Ser Ile Leu Leu Cys Leu Leu Ala Leu Arg Ala Glu Ala GlyPro 20 25 30 Pro Gln Glu Glu Ser Leu Tyr Leu Trp Ile Asp Ala His Gln Ala35 40 45 Arg Val Leu Ile Gly Phe Glu Glu Asp Ile Leu Ile Val Ser Glu 5055 60 Gly Lys Met Ala Pro Phe Thr His Asp Phe Arg Lys Ala Gln Gln 65 7075 Arg Met Pro Ala Ile Pro Val Asn Ile His Ser Met Asn Phe Thr 80 85 90Trp Gln Ala Ala Gly Gln Ala Glu Tyr Phe Tyr Glu Phe Leu Ser 95 100 105Leu Arg Ser Leu Asp Lys Gly Ile Met Ala Asp Pro Thr Val Asn 110 115 120Val Pro Leu Leu Gly Thr Val Pro His Lys Ala Ser Val Val Gln 125 130 135Val Gly Phe Pro Cys Leu Gly Lys Gln Asp Gly Val Ala Ala Phe 140 145 150Glu Val Asp Val Ile Val Met Asn Ser Glu Gly Asn Thr Ile Leu 155 160 165Gln Thr Pro Gln Asn Ala Ile Phe Phe Lys Thr Cys Gln Gln Ala 170 175 180Glu Cys Pro Gly Gly Cys Arg Asn Gly Gly Phe Cys Asn Glu Arg 185 190 195Arg Ile Cys Glu Cys Pro Asp Gly Phe His Gly Pro His Cys Glu 200 205 210Lys Ala Leu Cys Thr Pro Arg Cys Met Asn Gly Gly Leu Cys Val 215 220 225Thr Pro Gly Phe Cys Ile Cys Pro Pro Gly Phe Tyr Gly Val Asn 230 235 240Cys Asp Lys Ala Asn Cys Ser Thr Thr Cys Phe Asn Gly Gly Thr 245 250 255Cys Phe Tyr Pro Gly Lys Cys Ile Cys Pro Pro Gly Leu Glu Gly 260 265 270Glu Gln Cys Glu Ile Ser Lys Cys Pro Gln Pro Cys Arg Asn Gly 275 280 285Gly Lys Cys Ile Gly Lys Ser Lys Cys Lys Cys Ser Lys Gly Tyr 290 295 300Gln Gly Asp Leu Cys Ser Lys Pro Val Cys Glu Pro Gly Cys Gly 305 310 315Ala His Gly Thr Cys His Glu Pro Asn Lys Cys Gln Cys Gln Glu 320 325 330Gly Trp His Gly Arg His Cys Asn Lys Arg Tyr Glu Ala Ser Leu 335 340 345Ile His Ala Leu Arg Pro Ala Gly Ala Gln Leu Arg Gln His Thr 350 355 360Pro Ser Leu Lys Lys Ala Glu Glu Arg Arg Asp Pro Pro Glu Ser 365 370 375Asn Tyr Ile Trp 23 783 DNA Homo Sapien 23 agaacctcag aaatgtgagttatttgggaa tggctgtttg taaatgtcct 50 tacgtaagcc aagaggaggt cttgacttggggtcccaggg gtaccgcaga 100 tcccagggac tggagcagca ctagcaagct ctggaggatgagccaggagt 150 ctggaattga ggctgagcca aagaccccag ggccgtctca gtctcataaa200 aggggatcag gcaggaggag tttgggagaa acctgagaag ggcctgattt 250gcagcatcat gatgggcctc tccttggcct ctgctgtgct cctggcctcc 300 ctcctgagtctccaccttgg aactgccaca cgtgggagtg acatatccaa 350 gacctgctgc ttccaatacagccacaagcc ccttccctgg acctgggtgc 400 gaagctatga attcaccagt aacagctgctcccagcgggc tgtgatattc 450 actaccaaaa gaggcaagaa agtctgtacc catccaaggaaaaaatgggt 500 gcaaaaatac atttctttac tgaaaactcc gaaacaattg tgactcagct550 gaattttcat ccgaggacgc ttggaccccg ctcttggctc tgcagccctc 600tggggagcct gcggaatctt ttctgaaggc tacatggacc cgctggggag 650 gagagggtgtttcctcccag agttacttta ataaaggttg ttcatagagt 700 tgaaaaaaaa aaaaaaaaaaaaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 750 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaaaaa 783 24 94 PRT Homo Sapien 24 Met Met Gly Leu Ser Leu Ala Ser Ala ValLeu Leu Ala Ser Leu 1 5 10 15 Leu Ser Leu His Leu Gly Thr Ala Thr ArgGly Ser Asp Ile Ser 20 25 30 Lys Thr Cys Cys Phe Gln Tyr Ser His Lys ProLeu Pro Trp Thr 35 40 45 Trp Val Arg Ser Tyr Glu Phe Thr Ser Asn Ser CysSer Gln Arg 50 55 60 Ala Val Ile Phe Thr Thr Lys Arg Gly Lys Lys Val CysThr His 65 70 75 Pro Arg Lys Lys Trp Val Gln Lys Tyr Ile Ser Leu Leu LysThr 80 85 90 Pro Lys Gln Leu 25 23 DNA Artificial Sequence Syntheticoligonucleotide probe 25 ggatcaggca ggaggagttt ggg 23 26 23 DNAArtificial Sequence Synthetic oligonucleotide probe 26 ggatgggtacagactttctt gcc 23 27 50 DNA Artificial Sequence Syntheticoligonucleotide probe 27 atgatgggcc tctccttggc ctctgctgtg ctcctggcctccctcctgag 50 28 3552 DNA Homo Sapien 28 gcgagaacct ttgcacgcgcacaaactacg gggacgattt ctgattgatt 50 tttggcgctt tcgatccacc ctcctcccttctcatgggac tttggggaca 100 aagcgtcccg accgcctcga gcgctcgagc agggcgctatccaggagcca 150 ggacagcgtc gggaaccaga ccatggctcc tggaccccaa gatccttaag200 ttcgtcgtct tcatcgtcgc ggttctgctg ccggtccggg ttgactctgc 250caccatcccc cggcaggacg aagttcccca gcagacagtg gccccacagc 300 aacagaggcgcagcctcaag gaggaggagt gtccagcagg atctcataga 350 tcagaatata ctggagcctgtaacccgtgc acagagggtg tggattacac 400 cattgcttcc aacaatttgc cttcttgcctgctatgtaca gtttgtaaat 450 caggtcaaac aaataaaagt tcctgtacca cgaccagagacaccgtgtgt 500 cagtgtgaaa aaggaagctt ccaggataaa aactcccctg agatgtgccg550 gacgtgtaga acagggtgtc ccagagggat ggtcaaggtc agtaattgta 600cgccccggag tgacatcaag tgcaaaaatg aatcagctgc cagttccact 650 gggaaaaccccagcagcgga ggagacagtg accaccatcc tggggatgct 700 tgcctctccc tatcactaccttatcatcat agtggtttta gtcatcattt 750 tagctgtggt tgtggttggc ttttcatgtcggaagaaatt catttcttac 800 ctcaaaggca tctgctcagg tggtggagga ggtcccgaacgtgtgcacag 850 agtccttttc cggcggcgtt catgtccttc acgagttcct ggggcggagg900 acaatgcccg caacgagacc ctgagtaaca gatacttgca gcccacccag 950gtctctgagc aggaaatcca aggtcaggag ctggcagagc taacaggtgt 1000 gactgtagagtcgccagagg agccacagcg tctgctggaa caggcagaag 1050 ctgaagggtg tcagaggaggaggctgctgg ttccagtgaa tgacgctgac 1100 tccgctgaca tcagcacctt gctggatgcctcggcaacac tggaagaagg 1150 acatgcaaag gaaacaattc aggaccaact ggtgggctccgaaaagctct 1200 tttatgaaga agatgaggca ggctctgcta cgtcctgcct gtgaaagaat1250 ctcttcagga aaccagagct tccctcattt accttttctc ctacaaaggg 1300aagcagcctg gaagaaacag tccagtactt gacccatgcc ccaacaaact 1350 ctactatccaatatggggca gcttaccaat ggtcctagaa ctttgttaac 1400 gcacttggag taatttttatgaaatactgc gtgtgataag caaacgggag 1450 aaatttatat cagattcttg gctgcatagttatacgattg tgtattaagg 1500 gtcgttttag gccacatgcg gtggctcatg cctgtaatcccagcactttg 1550 ataggctgag gcaggtggat tgcttgagct cgggagtttg agaccagcct1600 catcaacaca gtgaaactcc atctcaattt aaaaagaaaa aaagtggttt 1650taggatgtca ttctttgcag ttcttcatca tgagacaagt ctttttttct 1700 gcttcttatattgcaagctc catctctact ggtgtgtgca tttaatgaca 1750 tctaactaca gatgccgcacagccacaatg ctttgcctta tagtttttta 1800 actttagaac gggattatct tgttattacctgtattttca gtttcggata 1850 tttttgactt aatgatgaga ttatcaagac gtagccctatgctaagtcat 1900 gagcatatgg acttacgagg gttcgactta gagttttgag ctttaagata1950 ggattattgg ggcttacccc caccttaatt agagaaacat ttatattgct 2000tactactgta ggctgtacat ctcttttccg atttttgtat aatgatgtaa 2050 acatggaaaaactttaggaa atgcacttat taggctgttt acatgggttg 2100 cctggataca aatcagcagtcaaaaatgac taaaaatata actagtgacg 2150 gagggagaaa tcctccctct gtgggaggcacttactgcat tccagttctc 2200 cctcctgcgc cctgagactg gaccagggtt tgatggctggcagcttctca 2250 aggggcagct tgtcttactt gttaatttta gaggtatata gccatattta2300 tttataaata aatatttatt tatttattta taagtagatg tttacatatg 2350cccaggattt tgaagagcct ggtatctttg ggaagccatg tgtctggttt 2400 gtcgtgctgggacagtcatg ggactgcatc ttccgacttg tccacagcag 2450 atgaggacag tgagaattaagttagatccg agactgcgaa gagcttctct 2500 ttcaagcgcc attacagttg aacgttagtgaatcttgagc ctcatttggg 2550 ctcagggcag agcaggtgtt tatctgcccc ggcatctgccatggcatcaa 2600 gagggaagag tggacggtgc ttgggaatgg tgtgaaatgg ttgccgactc2650 aggcatggat gggcccctct cgcttctggt ggtctgtgaa ctgagtccct 2700gggatgcctt ttagggcaga gattcctgag ctgcgtttta gggtacagat 2750 tccctgtttgaggagcttgg cccctctgta agcatctgac tcatctcaga 2800 gatatcaatt cttaaacactgtgacaacgg gatctaaaat ggctgacaca 2850 tttgtccttg tgtcacgttc cattattttatttaaaaacc tcagtaatcg 2900 ttttagcttc tttccagcaa actcttctcc acagtagcccagtcgtggta 2950 ggataaatta cggatatagt cattctaggg gtttcagtct tttccatctc3000 aaggcattgt gtgttttgtt ccgggactgg tttggctggg acaaagttag 3050aactgcctga agttcgcaca ttcagattgt tgtgtccatg gagttttagg 3100 aggggatggcctttccggtc ttcgcacttc catcctctcc cacttccatc 3150 tggcgtccca caccttgtcccctgcacttc tggatgacac agggtgctgc 3200 tgcctcctag tctttgcctt tgctgggccttctgtgcagg agacttggtc 3250 tcaaagctca gagagagcca gtccggtccc agctcctttgtcccttcctc 3300 agaggccttc cttgaagatg catctagact accagcctta tcagtgttta3350 agcttattcc tttaacataa gcttcctgac aacatgaaat tgttggggtt 3400ttttggcgtt ggttgatttg tttaggtttt gctttatacc cgggccaaat 3450 agcacataacacctggttat atatgaaata ctcatatgtt tatgaccaaa 3500 ataaatatga aacctcatrttaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 3550 aa 3552 29 386 PRT Homo Sapien 29Met Gly Leu Trp Gly Gln Ser Val Pro Thr Ala Ser Ser Ala Arg 1 5 10 15Ala Gly Arg Tyr Pro Gly Ala Arg Thr Ala Ser Gly Thr Arg Pro 20 25 30 TrpLeu Leu Asp Pro Lys Ile Leu Lys Phe Val Val Phe Ile Val 35 40 45 Ala ValLeu Leu Pro Val Arg Val Asp Ser Ala Thr Ile Pro Arg 50 55 60 Gln Asp GluVal Pro Gln Gln Thr Val Ala Pro Gln Gln Gln Arg 65 70 75 Arg Ser Leu LysGlu Glu Glu Cys Pro Ala Gly Ser His Arg Ser 80 85 90 Glu Tyr Thr Gly AlaCys Asn Pro Cys Thr Glu Gly Val Asp Tyr 95 100 105 Thr Ile Ala Ser AsnAsn Leu Pro Ser Cys Leu Leu Cys Thr Val 110 115 120 Cys Lys Ser Gly GlnThr Asn Lys Ser Ser Cys Thr Thr Thr Arg 125 130 135 Asp Thr Val Cys GlnCys Glu Lys Gly Ser Phe Gln Asp Lys Asn 140 145 150 Ser Pro Glu Met CysArg Thr Cys Arg Thr Gly Cys Pro Arg Gly 155 160 165 Met Val Lys Val SerAsn Cys Thr Pro Arg Ser Asp Ile Lys Cys 170 175 180 Lys Asn Glu Ser AlaAla Ser Ser Thr Gly Lys Thr Pro Ala Ala 185 190 195 Glu Glu Thr Val ThrThr Ile Leu Gly Met Leu Ala Ser Pro Tyr 200 205 210 His Tyr Leu Ile IleIle Val Val Leu Val Ile Ile Leu Ala Val 215 220 225 Val Val Val Gly PheSer Cys Arg Lys Lys Phe Ile Ser Tyr Leu 230 235 240 Lys Gly Ile Cys SerGly Gly Gly Gly Gly Pro Glu Arg Val His 245 250 255 Arg Val Leu Phe ArgArg Arg Ser Cys Pro Ser Arg Val Pro Gly 260 265 270 Ala Glu Asp Asn AlaArg Asn Glu Thr Leu Ser Asn Arg Tyr Leu 275 280 285 Gln Pro Thr Gln ValSer Glu Gln Glu Ile Gln Gly Gln Glu Leu 290 295 300 Ala Glu Leu Thr GlyVal Thr Val Glu Ser Pro Glu Glu Pro Gln 305 310 315 Arg Leu Leu Glu GlnAla Glu Ala Glu Gly Cys Gln Arg Arg Arg 320 325 330 Leu Leu Val Pro ValAsn Asp Ala Asp Ser Ala Asp Ile Ser Thr 335 340 345 Leu Leu Asp Ala SerAla Thr Leu Glu Glu Gly His Ala Lys Glu 350 355 360 Thr Ile Gln Asp GlnLeu Val Gly Ser Glu Lys Leu Phe Tyr Glu 365 370 375 Glu Asp Glu Ala GlySer Ala Thr Ser Cys Leu 380 385 30 50 DNA Artificial Sequence Syntheticoligonucleotide probe 30 cataaaagtt cctgcaccat gaccagagac acagtgtgtcagtgtaaaga 50 31 963 DNA Homo Sapien 31 gcggcacctg gaagatgcgc ccattggctggtggcctgct caaggtggtg 50 ttcgtggtct tcgcctcctt gtgtgcctgg tattcggggtacctgctcgc 100 agagctcatt ccagatgcac ccctgtccag tgctgcctat agcatccgca150 gcatcgggga gaggcctgtc ctcaaagctc cagtccccaa aaggcaaaaa 200tgtgaccact ggactccctg cccatctgac acctatgcct acaggttact 250 cagcggaggtggcagaagca agtacgccaa aatctgcttt gaggataacc 300 tacttatggg agaacagctgggaaatgttg ccagaggaat aaacattgcc 350 attgtcaact atgtaactgg gaatgtgacagcaacacgat gttttgatat 400 gtatgaaggc gataactctg gaccgatgac aaagtttattcagagtgctg 450 ctccaaaatc cctgctcttc atggtgacct atgacgacgg aagcacaaga500 ctgaataacg atgccaagaa tgccatagaa gcacttggaa gtaaagaaat 550caggaacatg aaattcaggt ctagctgggt atttattgca gcaaaaggct 600 tggaactcccttccgaaatt cagagagaaa agatcaacca ctctgatgct 650 aagaacaaca gatattctggctggcctgca gagatccaga tagaaggctg 700 catacccaaa gaacgaagct gacactgcagggtcctgagt aaatgtgttc 750 tgtataaaca aatgcagctg gaatcgctca agaatcttatttttctaaat 800 ccaacagccc atatttgatg agtattttgg gtttgttgta aaccaatgaa850 catttgctag ttgtatcaaa tcttggtacg cagtattttt ataccagtat 900tttatgtagt gaagatgtca attagcagga aactaaaatg aatggaaatt 950 cttaaaaaaaaaa 963 32 235 PRT Homo Sapien 32 Met Arg Pro Leu Ala Gly Gly Leu LeuLys Val Val Phe Val Val 1 5 10 15 Phe Ala Ser Leu Cys Ala Trp Tyr SerGly Tyr Leu Leu Ala Glu 20 25 30 Leu Ile Pro Asp Ala Pro Leu Ser Ser AlaAla Tyr Ser Ile Arg 35 40 45 Ser Ile Gly Glu Arg Pro Val Leu Lys Ala ProVal Pro Lys Arg 50 55 60 Gln Lys Cys Asp His Trp Thr Pro Cys Pro Ser AspThr Tyr Ala 65 70 75 Tyr Arg Leu Leu Ser Gly Gly Gly Arg Ser Lys Tyr AlaLys Ile 80 85 90 Cys Phe Glu Asp Asn Leu Leu Met Gly Glu Gln Leu Gly AsnVal 95 100 105 Ala Arg Gly Ile Asn Ile Ala Ile Val Asn Tyr Val Thr GlyAsn 110 115 120 Val Thr Ala Thr Arg Cys Phe Asp Met Tyr Glu Gly Asp AsnSer 125 130 135 Gly Pro Met Thr Lys Phe Ile Gln Ser Ala Ala Pro Lys SerLeu 140 145 150 Leu Phe Met Val Thr Tyr Asp Asp Gly Ser Thr Arg Leu AsnAsn 155 160 165 Asp Ala Lys Asn Ala Ile Glu Ala Leu Gly Ser Lys Glu IleArg 170 175 180 Asn Met Lys Phe Arg Ser Ser Trp Val Phe Ile Ala Ala LysGly 185 190 195 Leu Glu Leu Pro Ser Glu Ile Gln Arg Glu Lys Ile Asn HisSer 200 205 210 Asp Ala Lys Asn Asn Arg Tyr Ser Gly Trp Pro Ala Glu IleGln 215 220 225 Ile Glu Gly Cys Ile Pro Lys Glu Arg Ser 230 235 33 18DNA Artificial Sequence Synthetic oligonucleotide probe 33 ggctggcctgcagagatc 18 34 20 DNA Artificial Sequence Synthetic oligonucleotideprobe 34 aatgtgacca ctggactccc 20 35 18 DNA Artificial SequenceSynthetic oligonucleotide probe 35 aggcttggaa ctcccttc 18 36 24 DNAArtificial Sequence Synthetic oligonucleotide probe 36 aagattcttgagcgattcca gctg 24 37 47 DNA Artificial Sequence Syntheticoligonucleotide probe 37 aatccctgct cttcatggtg acctatgacg acggaagcacaagactg 47 38 1215 DNA Homo Sapien 38 ccggggaggg gagggcccgt cccgcccctccccgtctctc cccgcccctc 50 cccgtccctc ccgccgaagc tccgtcccgc ccgcgggccggctccgccct 100 cacctcccgg ccgcggctgc cctctgcccg ggttgtccaa gatggagggc150 gctccaccgg ggtcgctcgc cctccggctc ctgctgttcg tggcgctacc 200cgcctccggc tggctgacga cgggcgcccc cgagccgccg ccgctgtccg 250 gagccccacaggacggcatc agaattaatg taactacact gaaagatgat 300 ggggacatat ctaaacagcaggttgttctt aacataacct atgagagtgg 350 acaggtgtat gtaaatgact tacctgtaaatagtggtgta acccgaataa 400 gctgtcagac tttgatagtg aagaatgaaa atcttgaaaatttggaggaa 450 aaagaatatt ttggaattgt cagtgtaagg attttagttc atgagtggcc500 tatgacatct ggttccagtt tgcaactaat tgtcattcaa gaagaggtag 550tagagattga tggaaaacaa gttcagcaaa aggatgtcac tgaaattgat 600 attttagttaagaaccgggg agtactcaga cattcaaact ataccctccc 650 tttggaagaa agcatgctctactctatttc tcgagacagt gacattttat 700 ttacccttcc taacctctcc aaaaaagaaagtgttagttc actgcaaacc 750 actagccagt atcttatcag gaatgtggaa accactgtagatgaagatgt 800 tttacctggc aagttacctg aaactcctct cagagcagag ccgccatctt850 catataaggt aatgtgtcag tggatggaaa agtttagaaa agatctgtgt 900aggttctgga gcaacgtttt cccagtattc tttcagtttt tgaacatcat 950 ggtggttggaattacaggag cagctgtggt aataaccatc ttaaaggtgt 1000 ttttcccagt ttctgaatacaaaggaattc ttcagttgga taaagtggac 1050 gtcatacctg tgacagctat caacttatatccagatggtc cagagaaaag 1100 agctgaaaac cttgaagata aaacatgtat ttaaaacgccatctcatatc 1150 atggactccg aagtagcctg ttgcctccaa atttgccact tgaatataat1200 tttctttaaa tcgtt 1215 39 330 PRT Homo Sapien 39 Met Glu Gly Ala ProPro Gly Ser Leu Ala Leu Arg Leu Leu Leu 1 5 10 15 Phe Val Ala Leu ProAla Ser Gly Trp Leu Thr Thr Gly Ala Pro 20 25 30 Glu Pro Pro Pro Leu SerGly Ala Pro Gln Asp Gly Ile Arg Ile 35 40 45 Asn Val Thr Thr Leu Lys AspAsp Gly Asp Ile Ser Lys Gln Gln 50 55 60 Val Val Leu Asn Ile Thr Tyr GluSer Gly Gln Val Tyr Val Asn 65 70 75 Asp Leu Pro Val Asn Ser Gly Val ThrArg Ile Ser Cys Gln Thr 80 85 90 Leu Ile Val Lys Asn Glu Asn Leu Glu AsnLeu Glu Glu Lys Glu 95 100 105 Tyr Phe Gly Ile Val Ser Val Arg Ile LeuVal His Glu Trp Pro 110 115 120 Met Thr Ser Gly Ser Ser Leu Gln Leu IleVal Ile Gln Glu Glu 125 130 135 Val Val Glu Ile Asp Gly Lys Gln Val GlnGln Lys Asp Val Thr 140 145 150 Glu Ile Asp Ile Leu Val Lys Asn Arg GlyVal Leu Arg His Ser 155 160 165 Asn Tyr Thr Leu Pro Leu Glu Glu Ser MetLeu Tyr Ser Ile Ser 170 175 180 Arg Asp Ser Asp Ile Leu Phe Thr Leu ProAsn Leu Ser Lys Lys 185 190 195 Glu Ser Val Ser Ser Leu Gln Thr Thr SerGln Tyr Leu Ile Arg 200 205 210 Asn Val Glu Thr Thr Val Asp Glu Asp ValLeu Pro Gly Lys Leu 215 220 225 Pro Glu Thr Pro Leu Arg Ala Glu Pro ProSer Ser Tyr Lys Val 230 235 240 Met Cys Gln Trp Met Glu Lys Phe Arg LysAsp Leu Cys Arg Phe 245 250 255 Trp Ser Asn Val Phe Pro Val Phe Phe GlnPhe Leu Asn Ile Met 260 265 270 Val Val Gly Ile Thr Gly Ala Ala Val ValIle Thr Ile Leu Lys 275 280 285 Val Phe Phe Pro Val Ser Glu Tyr Lys GlyIle Leu Gln Leu Asp 290 295 300 Lys Val Asp Val Ile Pro Val Thr Ala IleAsn Leu Tyr Pro Asp 305 310 315 Gly Pro Glu Lys Arg Ala Glu Asn Leu GluAsp Lys Thr Cys Ile 320 325 330 40 2498 DNA Homo Sapien 40 cgtctctgcgttcgccatgc gtcccggggc gccagggcca ctctggcctc 50 tgccctgggg ggccctggcttgggccgtgg gcttcgtgag ctccatgggc 100 tcggggaacc ccgcgcccgg tggtgtttgctggctccagc agggccagga 150 ggccacctgc agcctggtgc tccagactga tgtcacccgggccgagtgct 200 gtgcctccgg caacattgac accgcctggt ccaacctcac ccacccgggg250 aacaagatca acctcctcgg cttcttgggc cttgtccact gccttccctg 300caaagattcg tgcgacggcg tggagtgcgg cccgggcaag gcgtgccgca 350 tgctggggggccgcccgcgc tgcgagtgcg cgcccgactg ctcggggctc 400 ccggcgcggc tgcaggtctgcggctcagac ggcgccacct accgcgacga 450 gtgcgagctg cgcgccgcgc gctgccgcggccacccggac ctgagcgtca 500 tgtaccgggg ccgctgccgc aagtcctgtg agcacgtggtgtgcccgcgg 550 ccacagtcgt gcgtcgtgga ccagacgggc agcgcccact gcgtggtgtg600 tcgagcggcg ccctgccctg tgccctccag ccccggccag gagctttgcg 650gcaacaacaa cgtcacctac atctcctcgt gccacatgcg ccaggccacc 700 tgcttcctgggccgctccat cggcgtgcgc cacgcgggca gctgcgcagg 750 cacccctgag gagccgccaggtggtgagtc tgcagaagag gaagagaact 800 tcgtgtgagc ctgcaggaca ggcctgggcctggtgcccga ggccccccat 850 catcccctgt tatttattgc cacagcagag tctaatttatatgccacgga 900 cactccttag agcccggatt cggaccactt ggggatccca gaacctccct950 gacgatatcc tggaaggact gaggaaggga ggcctggggg ccggctggtg 1000ggtgggatag acctgcgttc cggacactga gcgcctgatt tagggccctt 1050 ctctaggatgccccagcccc taccctaaga cctattgccg gggaggattc 1100 cacacttccg ctcctttggggataaaccta ttaattattg ctactatcaa 1150 gagggctggg cattctctgc tggtaattcctgaagaggca tgactgcttt 1200 tctcagcccc aagcctctag tctgggtgtg tacggagggtctagcctggg 1250 tgtgtacgga gggtctagcc tgggtgagta cggagggtct agcctgggtg1300 agtacggagg gtctagcctg ggtgagtacg gagggtctag cctgggtgtg 1350tatggaggat ctagcctggg tgagtatgga gggtctagcc tgggtgagta 1400 tggagggtctagcctgggtg tgtatggagg gtctagcctg ggtgagtatg 1450 gagggtctag cctgggtgtgtatggagggt ctagcctggg tgagtatgga 1500 gggtctagcc tgggtgtgta cggagggtctagtctgagtg cgtgtgggga 1550 cctcagaaca ctgtgacctt agcccagcaa gccaggcccttcatgaaggc 1600 caagaaggct gccaccattc cctgccagcc caagaactcc agcttcccca1650 ctgcctctgt gtgccccttt gcgtcctgtg aaggccattg agaaatgccc 1700agtgtgcccc ctgggaaagg gcacggcctg tgctcctgac acgggctgtg 1750 cttggccacagaaccaccca gcgtctcccc tgctgctgtc cacgtcagtt 1800 catgaggcaa cgtcgcgtggtctcagacgt ggagcagcca gcggcagctc 1850 agagcagggc actgtgtccg gcggagccaagtccactctg ggggagctct 1900 ggcggggacc acgggccact gctcacccac tggccccgaggggggtgtag 1950 acgccaagac tcacgcatgt gtgacatccg gagtcctgga gccgggtgtc2000 ccagtggcac cactaggtgc ctgctgcctc cacagtgggg ttcacaccca 2050gggctccttg gtcccccaca acctgccccg gccaggcctg cagacccaga 2100 ctccagccagacctgcctca cccaccaatg cagccggggc tggcgacacc 2150 agccaggtgc tggtcttgggccagttctcc cacgacggct caccctcccc 2200 tccatctgcg ttgatgctca gaatcgcctacctgtgcctg cgtgtaaacc 2250 acagcctcag accagctatg gggagaggac aacacggaggatatccagct 2300 tccccggtct ggggtgagga atgtggggag cttgggcatc ctcctccagc2350 ctcctccagc ccccaggcag tgccttacct gtggtgccca gaaaagtgcc 2400cctaggttgg tgggtctaca ggagcctcag ccaggcagcc caccccaccc 2450 tggggccctgcctcaccaag gaaataaaga ctcaagccat aaaaaaaa 2498 41 263 PRT Homo Sapien 41Met Arg Pro Gly Ala Pro Gly Pro Leu Trp Pro Leu Pro Trp Gly 1 5 10 15Ala Leu Ala Trp Ala Val Gly Phe Val Ser Ser Met Gly Ser Gly 20 25 30 AsnPro Ala Pro Gly Gly Val Cys Trp Leu Gln Gln Gly Gln Glu 35 40 45 Ala ThrCys Ser Leu Val Leu Gln Thr Asp Val Thr Arg Ala Glu 50 55 60 Cys Cys AlaSer Gly Asn Ile Asp Thr Ala Trp Ser Asn Leu Thr 65 70 75 His Pro Gly AsnLys Ile Asn Leu Leu Gly Phe Leu Gly Leu Val 80 85 90 His Cys Leu Pro CysLys Asp Ser Cys Asp Gly Val Glu Cys Gly 95 100 105 Pro Gly Lys Ala CysArg Met Leu Gly Gly Arg Pro Arg Cys Glu 110 115 120 Cys Ala Pro Asp CysSer Gly Leu Pro Ala Arg Leu Gln Val Cys 125 130 135 Gly Ser Asp Gly AlaThr Tyr Arg Asp Glu Cys Glu Leu Arg Ala 140 145 150 Ala Arg Cys Arg GlyHis Pro Asp Leu Ser Val Met Tyr Arg Gly 155 160 165 Arg Cys Arg Lys SerCys Glu His Val Val Cys Pro Arg Pro Gln 170 175 180 Ser Cys Val Val AspGln Thr Gly Ser Ala His Cys Val Val Cys 185 190 195 Arg Ala Ala Pro CysPro Val Pro Ser Ser Pro Gly Gln Glu Leu 200 205 210 Cys Gly Asn Asn AsnVal Thr Tyr Ile Ser Ser Cys His Met Arg 215 220 225 Gln Ala Thr Cys PheLeu Gly Arg Ser Ile Gly Val Arg His Ala 230 235 240 Gly Ser Cys Ala GlyThr Pro Glu Glu Pro Pro Gly Gly Glu Ser 245 250 255 Ala Glu Glu Glu GluAsn Phe Val 260 42 20 DNA Artificial Sequence Synthetic oligonucleotideprobe 42 tcctgtgagc acgtggtgtg 20 43 18 DNA Artificial SequenceSynthetic oligonucleotide probe 43 gggtgggata gacctgcg 18 44 18 DNAArtificial Sequence Synthetic oligonucleotide probe 44 aaggccaagaaggctgcc 18 45 18 DNA Artificial Sequence Synthetic oligonucleotideprobe 45 ccaggcctgc agacccag 18 46 24 DNA Artificial Sequence Syntheticoligonucleotide probe 46 cttcctcagt ccttccagga tatc 24 47 24 DNAArtificial Sequence Synthetic oligonucleotide probe 47 aagctggatatcctccgtgt tgtc 24 48 27 DNA Artificial Sequence Syntheticoligonucleotide probe 48 cctgaagagg catgactgct tttctca 27 49 27 DNAArtificial Sequence Synthetic oligonucleotide probe 49 ggggataaacctattaatta ttgctac 27 50 44 DNA Artificial Sequence Syntheticoligonucleotide probe 50 aacgtcacct acatctcctc gtgccacatg cgccaggccacctg 44 51 1690 DNA Homo Sapien 51 tgcagagctt gtggaggcca tggggcgcgtcgtcgcggag ctcgtctcct 50 cgctgctggg gttgtggctg ttgctgtgca gctgcggatgccccgagggc 100 gccgagctgc gtgctccgcc agataaaatc gcgattattg gagccggaat150 tggtggcact tcagcagcct attacctgcg gcagaaattt gggaaagatg 200tgaagataga cctgtttgaa agagaagagg tcgggggccg cctggctacc 250 atgatggtgcaggggcaaga atacgaggca ggaggttctg tcatccatcc 300 tttaaatctg cacatgaaacgttttgtcaa agacctgggt ctctctgctg 350 ttcaggcctc tggtggccta ctggggatatataatggaga gactctggta 400 tttgaggaga gcaactggtt cataattaac gtgattaaattagtttggcg 450 ctatggattt caatccctcc gtatgcacat gtgggtagag gacgtgttag500 acaagttcat gaggatctac cgctaccagt ctcatgacta tgccttcagt 550agtgtcgaaa aattacttca tgctctagga ggagatgact tccttggaat 600 gcttaatcgaacacttcttg aaaccttgca aaaggccggc ttttctgaga 650 agttcctcaa tgaaatgattgctcctgtta tgagggtcaa ttatggccaa 700 agcacggaca tcaatgcctt tgtgggggcggtgtcactgt cctgttctga 750 ttctggcctt tgggcagtag aaggtggcaa taaacttgtttgctcagggc 800 ttctgcaggc atccaaaagc aatcttatat ctggctcagt aatgtacatc850 gaggagaaaa caaagaccaa gtacacagga aatccaacaa agatgtatga 900agtggtctac caaattggaa ctgagactcg ttcagacttc tatgacatcg 950 tcttggtggccactccgttg aatcgaaaaa tgtcgaatat tacttttctc 1000 aactttgatc ctccaattgaggaattccat caatattatc aacatatagt 1050 gacaacttta gttaaggggg aattgaatacatctatcttt agctctagac 1100 ccatagataa atttggcctt aatacagttt taaccactgataattcagat 1150 ttgttcatta acagtattgg gattgtgccc tctgtgagag aaaaggaaga1200 tcctgagcca tcaacagatg gaacatatgt ttggaagatc ttttcccaag 1250aaactcttac taaagcacaa attttaaagc tctttctgtc ctatgattat 1300 gctgtgaagaagccatggct tgcatatcct cactataagc ccccggagaa 1350 atgcccctct atcattctccatgatcgact ttattacctc aatggcatag 1400 agtgtgcagc aagtgccatg gagatgagtgccattgcagc ccacaacgct 1450 gcactccttg cctatcaccg ctggaacggg cacacagacatgattgatca 1500 ggatggctta tatgagaaac ttaaaactga actatgaagt gacacactcc1550 tttttcccct cctagttcca aatgactatc agtggcaaaa aagaacaaaa 1600tctgagcaga gatgattttg aaccagatat tttgccatta tcattgttta 1650 ataaaagtaatccctgctgg tcataggaaa aaaaaaaaaa 1690 52 505 PRT Homo Sapien 52 Met GlyArg Val Val Ala Glu Leu Val Ser Ser Leu Leu Gly Leu 1 5 10 15 Trp LeuLeu Leu Cys Ser Cys Gly Cys Pro Glu Gly Ala Glu Leu 20 25 30 Arg Ala ProPro Asp Lys Ile Ala Ile Ile Gly Ala Gly Ile Gly 35 40 45 Gly Thr Ser AlaAla Tyr Tyr Leu Arg Gln Lys Phe Gly Lys Asp 50 55 60 Val Lys Ile Asp LeuPhe Glu Arg Glu Glu Val Gly Gly Arg Leu 65 70 75 Ala Thr Met Met Val GlnGly Gln Glu Tyr Glu Ala Gly Gly Ser 80 85 90 Val Ile His Pro Leu Asn LeuHis Met Lys Arg Phe Val Lys Asp 95 100 105 Leu Gly Leu Ser Ala Val GlnAla Ser Gly Gly Leu Leu Gly Ile 110 115 120 Tyr Asn Gly Glu Thr Leu ValPhe Glu Glu Ser Asn Trp Phe Ile 125 130 135 Ile Asn Val Ile Lys Leu ValTrp Arg Tyr Gly Phe Gln Ser Leu 140 145 150 Arg Met His Met Trp Val GluAsp Val Leu Asp Lys Phe Met Arg 155 160 165 Ile Tyr Arg Tyr Gln Ser HisAsp Tyr Ala Phe Ser Ser Val Glu 170 175 180 Lys Leu Leu His Ala Leu GlyGly Asp Asp Phe Leu Gly Met Leu 185 190 195 Asn Arg Thr Leu Leu Glu ThrLeu Gln Lys Ala Gly Phe Ser Glu 200 205 210 Lys Phe Leu Asn Glu Met IleAla Pro Val Met Arg Val Asn Tyr 215 220 225 Gly Gln Ser Thr Asp Ile AsnAla Phe Val Gly Ala Val Ser Leu 230 235 240 Ser Cys Ser Asp Ser Gly LeuTrp Ala Val Glu Gly Gly Asn Lys 245 250 255 Leu Val Cys Ser Gly Leu LeuGln Ala Ser Lys Ser Asn Leu Ile 260 265 270 Ser Gly Ser Val Met Tyr IleGlu Glu Lys Thr Lys Thr Lys Tyr 275 280 285 Thr Gly Asn Pro Thr Lys MetTyr Glu Val Val Tyr Gln Ile Gly 290 295 300 Thr Glu Thr Arg Ser Asp PheTyr Asp Ile Val Leu Val Ala Thr 305 310 315 Pro Leu Asn Arg Lys Met SerAsn Ile Thr Phe Leu Asn Phe Asp 320 325 330 Pro Pro Ile Glu Glu Phe HisGln Tyr Tyr Gln His Ile Val Thr 335 340 345 Thr Leu Val Lys Gly Glu LeuAsn Thr Ser Ile Phe Ser Ser Arg 350 355 360 Pro Ile Asp Lys Phe Gly LeuAsn Thr Val Leu Thr Thr Asp Asn 365 370 375 Ser Asp Leu Phe Ile Asn SerIle Gly Ile Val Pro Ser Val Arg 380 385 390 Glu Lys Glu Asp Pro Glu ProSer Thr Asp Gly Thr Tyr Val Trp 395 400 405 Lys Ile Phe Ser Gln Glu ThrLeu Thr Lys Ala Gln Ile Leu Lys 410 415 420 Leu Phe Leu Ser Tyr Asp TyrAla Val Lys Lys Pro Trp Leu Ala 425 430 435 Tyr Pro His Tyr Lys Pro ProGlu Lys Cys Pro Ser Ile Ile Leu 440 445 450 His Asp Arg Leu Tyr Tyr LeuAsn Gly Ile Glu Cys Ala Ala Ser 455 460 465 Ala Met Glu Met Ser Ala IleAla Ala His Asn Ala Ala Leu Leu 470 475 480 Ala Tyr His Arg Trp Asn GlyHis Thr Asp Met Ile Asp Gln Asp 485 490 495 Gly Leu Tyr Glu Lys Leu LysThr Glu Leu 500 505 53 728 DNA Homo Sapien 53 catttccaac aagagcactggccaagtcag cttcttctga gagagtctct 50 agaagacatg atgctacact cagctttgggtctctgcctc ttactcgtca 100 cagtttcttc caaccttgcc attgcaataa aaaaggaaaagaggcctcct 150 cagacactct caagaggatg gggagatgac atcacttggg tacaaactta200 tgaagaaggt ctcttttatg ctcaaaaaag taagaagcca ttaatggtta 250ttcatcacct ggaggattgt caatactctc aagcactaaa gaaagtattt 300 gcccaaaatgaagaaataca agaaatggct cagaataagt tcatcatgct 350 aaaccttatg catgaaaccactgataagaa tttatcacct gatgggcaat 400 atgtgcctag aatcatgttt gtagacccttctttaacagt tagagctgac 450 atagctggaa gatactctaa cagattgtac acatatgagcctcgggattt 500 acccctattg atagaaaaca tgaagaaagc attaagactt attcagtcag550 agctataaga gatgatggaa aaaagccttc acttcaaaga agtcaaattt 600catgaagaaa acctctggca cattgacaaa tactaaatgt gcaagtatat 650 agattttgtaatattactat ttagtttttt taatgtgttt gcaatagtct 700 tattaaaata aatgttttttaaatctga 728 54 166 PRT Homo Sapien 54 Met Met Leu His Ser Ala Leu GlyLeu Cys Leu Leu Leu Val Thr 1 5 10 15 Val Ser Ser Asn Leu Ala Ile AlaIle Lys Lys Glu Lys Arg Pro 20 25 30 Pro Gln Thr Leu Ser Arg Gly Trp GlyAsp Asp Ile Thr Trp Val 35 40 45 Gln Thr Tyr Glu Glu Gly Leu Phe Tyr AlaGln Lys Ser Lys Lys 50 55 60 Pro Leu Met Val Ile His His Leu Glu Asp CysGln Tyr Ser Gln 65 70 75 Ala Leu Lys Lys Val Phe Ala Gln Asn Glu Glu IleGln Glu Met 80 85 90 Ala Gln Asn Lys Phe Ile Met Leu Asn Leu Met His GluThr Thr 95 100 105 Asp Lys Asn Leu Ser Pro Asp Gly Gln Tyr Val Pro ArgIle Met 110 115 120 Phe Val Asp Pro Ser Leu Thr Val Arg Ala Asp Ile AlaGly Arg 125 130 135 Tyr Ser Asn Arg Leu Tyr Thr Tyr Glu Pro Arg Asp LeuPro Leu 140 145 150 Leu Ile Glu Asn Met Lys Lys Ala Leu Arg Leu Ile GlnSer Glu 155 160 165 Leu 55 537 DNA Homo Sapien 55 taaaacagct acaatattccagggccagtc acttgccatt tctcataaca 50 gcgtcagaga gaaagaactg actgaaacgtttgagatgaa gaaagttctc 100 ctcctgatca cagccatctt ggcagtggct gttggtttcccagtctctca 150 agaccaggaa cgagaaaaaa gaagtatcag tgacagcgat gaattagctt200 cagggttttt tgtgttccct tacccatatc catttcgccc acttccacca 250attccatttc caagatttcc atggtttaga cgtaattttc ctattccaat 300 acctgaatctgcccctacaa ctccccttcc tagcgaaaag taaacaagaa 350 ggataagtca cgataaacctggtcacctga aattgaaatt gagccacttc 400 cttgaagaat caaaattcct gttaataaaagaaaaacaaa tgtaattgaa 450 atagcacaca gcattctcta gtcaatatct ttagtgatcttctttaataa 500 acatgaaagc aaagattttg gtttcttaat ttccaca 537 56 85 PRTHomo Sapien 56 Met Lys Lys Val Leu Leu Leu Ile Thr Ala Ile Leu Ala ValAla 1 5 10 15 Val Gly Phe Pro Val Ser Gln Asp Gln Glu Arg Glu Lys ArgSer 20 25 30 Ile Ser Asp Ser Asp Glu Leu Ala Ser Gly Phe Phe Val Phe Pro35 40 45 Tyr Pro Tyr Pro Phe Arg Pro Leu Pro Pro Ile Pro Phe Pro Arg 5055 60 Phe Pro Trp Phe Arg Arg Asn Phe Pro Ile Pro Ile Pro Glu Ser 65 7075 Ala Pro Thr Thr Pro Leu Pro Ser Glu Lys 80 85 57 2997 DNA Homo Sapien57 cggacgcgtg ggcgggcgcg ccgggaggga ccggcggcgg catgggccgg 50 gggccctgggatgcgggccc gtctcgccgc ctgctgccgc tgttgctgct 100 gctcggcctg gcccgcggcgccgcgggagc gccgggcccc gacggtttag 150 acgtctgtgc cacttgccat gaacatgccacatgccagca aagagaaggg 200 aagaagatct gtatttgcaa ctatggattt gtagggaacgggaggactca 250 gtgtgttgat aaaaatgagt gccagtttgg agccactctt gtctgtggga300 accacacatc ttgccacaac acccccgggg gcttctattg catttgcctg 350gaaggatatc gagccacaaa caacaacaag acattcattc ccaacgatgg 400 caccttttgtacagacatag atgagtgtga agtttctggc ctgtgcaggc 450 atggagggcg atgcgtgaacactcatggga gctttgaatg ctactgtatg 500 gatggatact tgccaaggaa tggacctgaacctttccacc cgaccaccga 550 tgccacatca tgcacagaaa tagactgtgg tacccctcctgaggttccag 600 atggctatat cataggaaat tatacgtcta gtctgggcag ccaggttcgt650 tatgcttgca gagaaggatt cttcagtgtt ccagaagata cagtttcaag 700ctgcacaggc ctgggcacat gggagtcccc aaaattacat tgccaagaga 750 tcaactgtggcaaccctcca gaaatgcggc acgccatctt ggtaggaaat 800 cacagctcca ggctgggcggtgtggctcgc tatgtctgtc aagagggctt 850 tgagagccct ggaggaaaga tcacttctgtttgcacagag aaaggcacct 900 ggagagaaag tactttaaca tgcacagaaa ttctgacaaagattaatgat 950 gtatcactgt ttaatgatac ctgtgtgaga tggcaaataa actcaagaag1000 aataaacccc aagatctcat atgtgatatc cataaaagga caacggttgg 1050accctatgga atcagttcgt gaggagacag tcaacttgac cacagacagc 1100 aggaccccagaagtgtgcct agccctgtac ccaggcacca actacaccgt 1150 gaacatctcc acagcacctcccaggcgctc gatgccagcc gtcatcggtt 1200 tccagacagc tgaagttgat ctcttagaagatgatggaag tttcaatatt 1250 tcaatattta atgaaacttg tttgaaattg aacaggcgttctaggaaagt 1300 tggatcagaa cacatgtacc aatttaccgt tctgggtcag aggtggtatc1350 tggctaactt ttctcatgca acatcgttta acttcacaac gagggaacaa 1400gtgcctgtag tgtgtttgga tctgtaccct acgactgatt atacggtgaa 1450 tgtgaccctgctgagatctc ctaagcggca ctcagtgcaa ataacaatag 1500 caactccccc agcagtaaaacagaccatca gtaacatttc aggatttaat 1550 gaaacctgct tgagatggag aagcatcaagacagctgata tggaggagat 1600 gtatttattc cacatttggg gccagagatg gtatcagaaggaatttgccc 1650 aggaaatgac ctttaatatc agtagcagca gccgagatcc cgaggtgtgc1700 ttggacctac gtccgggtac caactacaat gtcagtctcc gggctctgtc 1750ttcggaactt cctgtggtca tctccctgac aacccagata acagagcctc 1800 ccctcccggaagtagaattt tttacggtgc acagaggacc tctaccacgc 1850 ctcagactga ggaaagccaaggagaaaaat ggaccaatca gttcatatca 1900 ggtgttagtg cttcccctgg ccctccaaagcacattttct tgtgattctg 1950 aaggcgcttc ctccttcttt agcaacgcct ctgatgctgatggatacgtg 2000 gctgcagaac tactggccaa agatgttcca gatgatgcca tggagatacc2050 tataggagac aggctgtact atggggaata ttataatgca cccttgaaaa 2100gagggagtga ttactgcatt atattacgaa tcacaagtga atggaataag 2150 gtgagaagacactcctgtgc agtttgggct caggtgaaag attcgtcact 2200 catgctgctg cagatggcgggtgttggact gggttccctg gctgttgtga 2250 tcattctcac attcctctcc ttctcagcggtgtgatggca gatggacact 2300 gagtggggag gatgcactgc tgctgggcag gtgttctggcagcttctcag 2350 gtgcccgcac agaggctccg tgtgacttcc gtccagggag catgtgggcc2400 tgcaactttc tccattccca gctgggcccc attcctggat ttaagatggt 2450ggctatccct gaggagtcac cataaggaga aaactcagga attctgagtc 2500 ttccctgctacaggaccagt tctgtgcaat gaacttgaga ctcctgatgt 2550 acactgtgat attgaccgaaggctacatac agatctgtga atcttggctg 2600 ggacttcctc tgagtgatgc ctgagggtcagctcctctag acattgactg 2650 caagagaatc tctgcaacct cctatataaa agcatttctgttaattcatt 2700 cagaatccat tctttacaat atgcagtgag atgggcttaa gtttgggcta2750 gagtttgact ttatgaagga ggtcattgaa aaagagaaca gtgacgtagg 2800caaatgtttc aagcacttta gaaacagtac ttttcctata attagttgat 2850 atactaatgagaaaatatac tagcctggcc atgccaataa gtttcctgct 2900 gtgtctgtta ggcagcattgctttgatgca atttctattg tcctatatat 2950 tcaaaagtaa tgtctacatt ccagtaaaaatatcccgtaa ttaaaaa 2997 58 747 PRT Homo Sapien 58 Met Gly Arg Gly ProTrp Asp Ala Gly Pro Ser Arg Arg Leu Leu 1 5 10 15 Pro Leu Leu Leu LeuLeu Gly Leu Ala Arg Gly Ala Ala Gly Ala 20 25 30 Pro Gly Pro Asp Gly LeuAsp Val Cys Ala Thr Cys His Glu His 35 40 45 Ala Thr Cys Gln Gln Arg GluGly Lys Lys Ile Cys Ile Cys Asn 50 55 60 Tyr Gly Phe Val Gly Asn Gly ArgThr Gln Cys Val Asp Lys Asn 65 70 75 Glu Cys Gln Phe Gly Ala Thr Leu ValCys Gly Asn His Thr Ser 80 85 90 Cys His Asn Thr Pro Gly Gly Phe Tyr CysIle Cys Leu Glu Gly 95 100 105 Tyr Arg Ala Thr Asn Asn Asn Lys Thr PheIle Pro Asn Asp Gly 110 115 120 Thr Phe Cys Thr Asp Ile Asp Glu Cys GluVal Ser Gly Leu Cys 125 130 135 Arg His Gly Gly Arg Cys Val Asn Thr HisGly Ser Phe Glu Cys 140 145 150 Tyr Cys Met Asp Gly Tyr Leu Pro Arg AsnGly Pro Glu Pro Phe 155 160 165 His Pro Thr Thr Asp Ala Thr Ser Cys ThrGlu Ile Asp Cys Gly 170 175 180 Thr Pro Pro Glu Val Pro Asp Gly Tyr IleIle Gly Asn Tyr Thr 185 190 195 Ser Ser Leu Gly Ser Gln Val Arg Tyr AlaCys Arg Glu Gly Phe 200 205 210 Phe Ser Val Pro Glu Asp Thr Val Ser SerCys Thr Gly Leu Gly 215 220 225 Thr Trp Glu Ser Pro Lys Leu His Cys GlnGlu Ile Asn Cys Gly 230 235 240 Asn Pro Pro Glu Met Arg His Ala Ile LeuVal Gly Asn His Ser 245 250 255 Ser Arg Leu Gly Gly Val Ala Arg Tyr ValCys Gln Glu Gly Phe 260 265 270 Glu Ser Pro Gly Gly Lys Ile Thr Ser ValCys Thr Glu Lys Gly 275 280 285 Thr Trp Arg Glu Ser Thr Leu Thr Cys ThrGlu Ile Leu Thr Lys 290 295 300 Ile Asn Asp Val Ser Leu Phe Asn Asp ThrCys Val Arg Trp Gln 305 310 315 Ile Asn Ser Arg Arg Ile Asn Pro Lys IleSer Tyr Val Ile Ser 320 325 330 Ile Lys Gly Gln Arg Leu Asp Pro Met GluSer Val Arg Glu Glu 335 340 345 Thr Val Asn Leu Thr Thr Asp Ser Arg ThrPro Glu Val Cys Leu 350 355 360 Ala Leu Tyr Pro Gly Thr Asn Tyr Thr ValAsn Ile Ser Thr Ala 365 370 375 Pro Pro Arg Arg Ser Met Pro Ala Val IleGly Phe Gln Thr Ala 380 385 390 Glu Val Asp Leu Leu Glu Asp Asp Gly SerPhe Asn Ile Ser Ile 395 400 405 Phe Asn Glu Thr Cys Leu Lys Leu Asn ArgArg Ser Arg Lys Val 410 415 420 Gly Ser Glu His Met Tyr Gln Phe Thr ValLeu Gly Gln Arg Trp 425 430 435 Tyr Leu Ala Asn Phe Ser His Ala Thr SerPhe Asn Phe Thr Thr 440 445 450 Arg Glu Gln Val Pro Val Val Cys Leu AspLeu Tyr Pro Thr Thr 455 460 465 Asp Tyr Thr Val Asn Val Thr Leu Leu ArgSer Pro Lys Arg His 470 475 480 Ser Val Gln Ile Thr Ile Ala Thr Pro ProAla Val Lys Gln Thr 485 490 495 Ile Ser Asn Ile Ser Gly Phe Asn Glu ThrCys Leu Arg Trp Arg 500 505 510 Ser Ile Lys Thr Ala Asp Met Glu Glu MetTyr Leu Phe His Ile 515 520 525 Trp Gly Gln Arg Trp Tyr Gln Lys Glu PheAla Gln Glu Met Thr 530 535 540 Phe Asn Ile Ser Ser Ser Ser Arg Asp ProGlu Val Cys Leu Asp 545 550 555 Leu Arg Pro Gly Thr Asn Tyr Asn Val SerLeu Arg Ala Leu Ser 560 565 570 Ser Glu Leu Pro Val Val Ile Ser Leu ThrThr Gln Ile Thr Glu 575 580 585 Pro Pro Leu Pro Glu Val Glu Phe Phe ThrVal His Arg Gly Pro 590 595 600 Leu Pro Arg Leu Arg Leu Arg Lys Ala LysGlu Lys Asn Gly Pro 605 610 615 Ile Ser Ser Tyr Gln Val Leu Val Leu ProLeu Ala Leu Gln Ser 620 625 630 Thr Phe Ser Cys Asp Ser Glu Gly Ala SerSer Phe Phe Ser Asn 635 640 645 Ala Ser Asp Ala Asp Gly Tyr Val Ala AlaGlu Leu Leu Ala Lys 650 655 660 Asp Val Pro Asp Asp Ala Met Glu Ile ProIle Gly Asp Arg Leu 665 670 675 Tyr Tyr Gly Glu Tyr Tyr Asn Ala Pro LeuLys Arg Gly Ser Asp 680 685 690 Tyr Cys Ile Ile Leu Arg Ile Thr Ser GluTrp Asn Lys Val Arg 695 700 705 Arg His Ser Cys Ala Val Trp Ala Gln ValLys Asp Ser Ser Leu 710 715 720 Met Leu Leu Gln Met Ala Gly Val Gly LeuGly Ser Leu Ala Val 725 730 735 Val Ile Ile Leu Thr Phe Leu Ser Phe SerAla Val 740 745 59 22 DNA Artificial Sequence Synthetic oligonucleotideprobe 59 ccacttgcca tgaacatgcc ac 22 60 25 DNA Artificial SequenceSynthetic oligonucleotide probe 60 cctcttgaca gacatagcga gccac 25 61 43DNA Artificial Sequence Synthetic oligonucleotide probe 61 cactcttgtctgtgggaacc acacatcttg ccacaactgt ggc 43 62 2015 DNA Homo Sapien 62ggaaaaggta cccgcgagag acagccagca gttctgtgga gcagcggtgg 50 ccggctaggatgggctgtct ctggggtctg gctctgcccc ttttcttctt 100 ctgctgggag gttggggtctctgggagctc tgcaggcccc agcacccgca 150 gagcagacac tgcgatgaca acggacgacacagaagtgcc cgctatgact 200 ctagcaccgg gccacgccgc tctggaaact caaacgctgagcgctgagac 250 ctcttctagg gcctcaaccc cagccggccc cattccagaa gcagagacca300 ggggagccaa gagaatttcc cctgcaagag agaccaggag tttcacaaaa 350acatctccca acttcatggt gctgatcgcc acctccgtgg agacatcagc 400 cgccagtggcagccccgagg gagctggaat gaccacagtt cagaccatca 450 caggcagtga tcccgaggaagccatctttg acaccctttg caccgatgac 500 agctctgaag aggcaaagac actcacaatggacatattga cattggctca 550 cacctccaca gaagctaagg gcctgtcctc agagagcagtgcctcttccg 600 acggccccca tccagtcatc accccgtcac gggcctcaga gagcagcgcc650 tcttccgacg gcccccatcc agtcatcacc ccgtcacggg cctcagagag 700cagcgcctct tccgacggcc cccatccagt catcaccccg tcatggtccc 750 cgggatctgatgtcactctc ctcgctgaag ccctggtgac tgtcacaaac 800 atcgaggtta ttaattgcagcatcacagaa atagaaacaa caacttccag 850 catccctggg gcctcagaca tagatctcatccccacggaa ggggtgaagg 900 cctcgtccac ctccgatcca ccagctctgc ctgactccactgaagcaaaa 950 ccacacatca ctgaggtcac agcctctgcc gagaccctgt ccacagccgg1000 caccacagag tcagctgcac ctcatgccac ggttgggacc ccactcccca 1050ctaacagcgc cacagaaaga gaagtgacag cacccggggc cacgaccctc 1100 agtggagctctggtcacagt tagcaggaat cccctggaag aaacctcagc 1150 cctctctgtt gagacaccaagttacgtcaa agtctcagga gcagctccgg 1200 tctccataga ggctgggtca gcagtgggcaaaacaacttc ctttgctggg 1250 agctctgctt cctcctacag cccctcggaa gccgccctcaagaacttcac 1300 cccttcagag acaccgacca tggacatcgc aaccaagggg cccttcccca1350 ccagcaggga ccctcttcct tctgtccctc cgactacaac caacagcagc 1400cgagggacga acagcacctt agccaagatc acaacctcag cgaagaccac 1450 gatgaagccccaacagccac gcccacgact gcccggacga ggccgaccac 1500 agacgtgagt gcaggtgaaaatggaggttt cctcctcctg cggctgagtg 1550 tggcttcccc ggaagacctc actgaccccagagtggcaga aaggctgatg 1600 cagcagctcc accgggaact ccacgcccac gcgcctcacttccaggtctc 1650 cttactgcgt gtcaggagag gctaacggac atcagctgca gccaggcatg1700 tcccgtatgc caaaagaggg tgctgcccct agcctgggcc cccaccgaca 1750gactgcagct gcgttactgt gctgagaggt acccagaagg ttcccatgaa 1800 gggcagcatgtccaagcccc taaccccaga tgtggcaaca ggaccctcgc 1850 tcacatccac cggagtgtatgtatggggag gggcttcacc tgttcccaga 1900 ggtgtccttg gactcacctt ggcacatgttctgtgtttca gtaaagagag 1950 acctgatcac ccatctgtgt gcttccatcc tgcattaaaattcactcagt 2000 gtggcccaaa aaaaa 2015 63 482 PRT Homo Sapien 63 Met GlyCys Leu Trp Gly Leu Ala Leu Pro Leu Phe Phe Phe Cys 1 5 10 15 Trp GluVal Gly Val Ser Gly Ser Ser Ala Gly Pro Ser Thr Arg 20 25 30 Arg Ala AspThr Ala Met Thr Thr Asp Asp Thr Glu Val Pro Ala 35 40 45 Met Thr Leu AlaPro Gly His Ala Ala Leu Glu Thr Gln Thr Leu 50 55 60 Ser Ala Glu Thr SerSer Arg Ala Ser Thr Pro Ala Gly Pro Ile 65 70 75 Pro Glu Ala Glu Thr ArgGly Ala Lys Arg Ile Ser Pro Ala Arg 80 85 90 Glu Thr Arg Ser Phe Thr LysThr Ser Pro Asn Phe Met Val Leu 95 100 105 Ile Ala Thr Ser Val Glu ThrSer Ala Ala Ser Gly Ser Pro Glu 110 115 120 Gly Ala Gly Met Thr Thr ValGln Thr Ile Thr Gly Ser Asp Pro 125 130 135 Glu Glu Ala Ile Phe Asp ThrLeu Cys Thr Asp Asp Ser Ser Glu 140 145 150 Glu Ala Lys Thr Leu Thr MetAsp Ile Leu Thr Leu Ala His Thr 155 160 165 Ser Thr Glu Ala Lys Gly LeuSer Ser Glu Ser Ser Ala Ser Ser 170 175 180 Asp Gly Pro His Pro Val IleThr Pro Ser Arg Ala Ser Glu Ser 185 190 195 Ser Ala Ser Ser Asp Gly ProHis Pro Val Ile Thr Pro Ser Arg 200 205 210 Ala Ser Glu Ser Ser Ala SerSer Asp Gly Pro His Pro Val Ile 215 220 225 Thr Pro Ser Trp Ser Pro GlySer Asp Val Thr Leu Leu Ala Glu 230 235 240 Ala Leu Val Thr Val Thr AsnIle Glu Val Ile Asn Cys Ser Ile 245 250 255 Thr Glu Ile Glu Thr Thr ThrSer Ser Ile Pro Gly Ala Ser Asp 260 265 270 Ile Asp Leu Ile Pro Thr GluGly Val Lys Ala Ser Ser Thr Ser 275 280 285 Asp Pro Pro Ala Leu Pro AspSer Thr Glu Ala Lys Pro His Ile 290 295 300 Thr Glu Val Thr Ala Ser AlaGlu Thr Leu Ser Thr Ala Gly Thr 305 310 315 Thr Glu Ser Ala Ala Pro HisAla Thr Val Gly Thr Pro Leu Pro 320 325 330 Thr Asn Ser Ala Thr Glu ArgGlu Val Thr Ala Pro Gly Ala Thr 335 340 345 Thr Leu Ser Gly Ala Leu ValThr Val Ser Arg Asn Pro Leu Glu 350 355 360 Glu Thr Ser Ala Leu Ser ValGlu Thr Pro Ser Tyr Val Lys Val 365 370 375 Ser Gly Ala Ala Pro Val SerIle Glu Ala Gly Ser Ala Val Gly 380 385 390 Lys Thr Thr Ser Phe Ala GlySer Ser Ala Ser Ser Tyr Ser Pro 395 400 405 Ser Glu Ala Ala Leu Lys AsnPhe Thr Pro Ser Glu Thr Pro Thr 410 415 420 Met Asp Ile Ala Thr Lys GlyPro Phe Pro Thr Ser Arg Asp Pro 425 430 435 Leu Pro Ser Val Pro Pro ThrThr Thr Asn Ser Ser Arg Gly Thr 440 445 450 Asn Ser Thr Leu Ala Lys IleThr Thr Ser Ala Lys Thr Thr Met 455 460 465 Lys Pro Gln Gln Pro Arg ProArg Leu Pro Gly Arg Gly Arg Pro 470 475 480 Gln Thr 64 1252 DNA HomoSapien 64 gcctctgaat tgttgggcag tctggcagtg gagctctccc cggtctgaca 50gccactccag aggccatgct tcgtttcttg ccagatttgg ctttcagctt 100 cctgttaattctggctttgg gccaggcagt ccaatttcaa gaatatgtct 150 ttctccaatt tctgggcttagataaggcgc cttcacccca gaagttccaa 200 cctgtgcctt atatcttgaa gaaaattttccaggatcgcg aggcagcagc 250 gaccactggg gtctcccgag acttatgcta cgtaaaggagctgggcgtcc 300 gcgggaatgt acttcgcttt ctcccagacc aaggtttctt tctttaccca350 aagaaaattt cccaagcttc ctcctgcctg cagaagctcc tctactttaa 400cctgtctgcc atcaaagaaa gggaacagtt gacattggcc cagctgggcc 450 tggacttggggcccaattct tactataacc tgggaccaga gctggaactg 500 gctctgttcc tggttcaggagcctcatgtg tggggccaga ccacccctaa 550 gccaggtaaa atgtttgtgt tgcggtcagtcccatggcca caaggtgctg 600 ttcacttcaa cctgctggat gtagctaagg attggaatgacaacccccgg 650 aaaaatttcg ggttattcct ggagatactg gtcaaagaag atagagactc700 aggggtgaat tttcagcctg aagacacctg tgccagacta agatgctccc 750ttcatgcttc cctgctggtg gtgactctca accctgatca gtgccaccct 800 tctcggaaaaggagagcagc catccctgtc cccaagcttt cttgtaagaa 850 cctctgccac cgtcaccagctattcattaa cttccgggac ctgggttggc 900 acaagtggat cattgccccc aaggggttcatggcaaatta ctgccatgga 950 gagtgtccct tctcactgac catctctctc aacagctccaattatgcttt 1000 catgcaagcc ctgatgcatg ccgttgaccc agagatcccc caggctgtgt1050 gtatccccac caagctgtct cccatttcca tgctctacca ggacaataat 1100gacaatgtca ttctacgaca ttatgaagac atggtagtcg atgaatgtgg 1150 gtgtgggtaggatgtcagaa atgggaatag aaggagtgtt cttagggtaa 1200 atcttttaat aaaactacctatctggttta tgaccactta gatcgaaatg 1250 tc 1252 65 364 PRT Homo Sapien 65Met Leu Arg Phe Leu Pro Asp Leu Ala Phe Ser Phe Leu Leu Ile 1 5 10 15Leu Ala Leu Gly Gln Ala Val Gln Phe Gln Glu Tyr Val Phe Leu 20 25 30 GlnPhe Leu Gly Leu Asp Lys Ala Pro Ser Pro Gln Lys Phe Gln 35 40 45 Pro ValPro Tyr Ile Leu Lys Lys Ile Phe Gln Asp Arg Glu Ala 50 55 60 Ala Ala ThrThr Gly Val Ser Arg Asp Leu Cys Tyr Val Lys Glu 65 70 75 Leu Gly Val ArgGly Asn Val Leu Arg Phe Leu Pro Asp Gln Gly 80 85 90 Phe Phe Leu Tyr ProLys Lys Ile Ser Gln Ala Ser Ser Cys Leu 95 100 105 Gln Lys Leu Leu TyrPhe Asn Leu Ser Ala Ile Lys Glu Arg Glu 110 115 120 Gln Leu Thr Leu AlaGln Leu Gly Leu Asp Leu Gly Pro Asn Ser 125 130 135 Tyr Tyr Asn Leu GlyPro Glu Leu Glu Leu Ala Leu Phe Leu Val 140 145 150 Gln Glu Pro His ValTrp Gly Gln Thr Thr Pro Lys Pro Gly Lys 155 160 165 Met Phe Val Leu ArgSer Val Pro Trp Pro Gln Gly Ala Val His 170 175 180 Phe Asn Leu Leu AspVal Ala Lys Asp Trp Asn Asp Asn Pro Arg 185 190 195 Lys Asn Phe Gly LeuPhe Leu Glu Ile Leu Val Lys Glu Asp Arg 200 205 210 Asp Ser Gly Val AsnPhe Gln Pro Glu Asp Thr Cys Ala Arg Leu 215 220 225 Arg Cys Ser Leu HisAla Ser Leu Leu Val Val Thr Leu Asn Pro 230 235 240 Asp Gln Cys His ProSer Arg Lys Arg Arg Ala Ala Ile Pro Val 245 250 255 Pro Lys Leu Ser CysLys Asn Leu Cys His Arg His Gln Leu Phe 260 265 270 Ile Asn Phe Arg AspLeu Gly Trp His Lys Trp Ile Ile Ala Pro 275 280 285 Lys Gly Phe Met AlaAsn Tyr Cys His Gly Glu Cys Pro Phe Ser 290 295 300 Leu Thr Ile Ser LeuAsn Ser Ser Asn Tyr Ala Phe Met Gln Ala 305 310 315 Leu Met His Ala ValAsp Pro Glu Ile Pro Gln Ala Val Cys Ile 320 325 330 Pro Thr Lys Leu SerPro Ile Ser Met Leu Tyr Gln Asp Asn Asn 335 340 345 Asp Asn Val Ile LeuArg His Tyr Glu Asp Met Val Val Asp Glu 350 355 360 Cys Gly Cys Gly 6620 DNA Artificial Sequence Synthetic oligonucleotide probe 66 gtctgacagccactccagag 20 67 47 DNA Artificial Sequence Synthetic oligonucleotideprobe 67 tctccaattt ctgggcttag ataaggcgcc ttcaccccag aagttcc 47 68 24DNA Artificial Sequence Synthetic oligonucleotide probe 68 gtcccaggttatagtaagaa ttgg 24 69 20 DNA Artificial Sequence Syntheticoligonucleotide probe 69 gtgttgcggt cagtcccatg 20 70 20 DNA ArtificialSequence Synthetic oligonucleotide probe 70 gctgtctccc atttccatgc 20 7124 DNA Artificial Sequence Synthetic oligonucleotide probe 71 cgactaccatgtcttcataa tgtc 24 72 2849 DNA Homo Sapien 72 cactttctcc ctctcttcctttactttcga gaaaccgcgc ttccgcttct 50 ggtcgcagag acctcggaga ccgcgccggggagacggagg tgctgtgggt 100 gggggggacc tgtggctgct cgtaccgccc cccaccctcctcttctgcac 150 tgccgtcctc cggaagacct tttcccctgc tctgtttcct tcaccgagtc200 tgtgcatcgc cccggacctg gccgggagga ggcttggccg gcgggagatg 250ctctaggggc ggcgcgggag gagcggccgg cgggacggag ggcccggcag 300 gaagatgggctcccgtggac agggactctt gctggcgtac tgcctgctcc 350 ttgcctttgc ctctggcctggtcctgagtc gtgtgcccca tgtccagggg 400 gaacagcagg agtgggaggg gactgaggagctgccgtcgc ctccggacca 450 tgccgagagg gctgaagaac aacatgaaaa atacaggcccagtcaggacc 500 aggggctccc tgcttcccgg tgcttgcgct gctgtgaccc cggtacctcc550 atgtacccgg cgaccgccgt gccccagatc aacatcacta tcttgaaagg 600ggagaagggt gaccgcggag atcgaggcct ccaagggaaa tatggcaaaa 650 caggctcagcaggggccagg ggccacactg gacccaaagg gcagaagggc 700 tccatggggg cccctggggagcggtgcaag agccactacg ccgccttttc 750 ggtgggccgg aagaagccca tgcacagcaaccactactac cagacggtga 800 tcttcgacac ggagttcgtg aacctctacg accacttcaacatgttcacc 850 ggcaagttct actgctacgt gcccggcctc tacttcttca gcctcaacgt900 gcacacctgg aaccagaagg agacctacct gcacatcatg aagaacgagg 950aggaggtggt gatcttgttc gcgcaggtgg gcgaccgcag catcatgcaa 1000 agccagagcctgatgctgga gctgcgagag caggaccagg tgtgggtacg 1050 cctctacaag ggcgaacgtgagaacgccat cttcagcgag gagctggaca 1100 cctacatcac cttcagtggc tacctggtcaagcacgccac cgagccctag 1150 ctggccggcc acctcctttc ctctcgccac cttccacccctgcgctgtgc 1200 tgaccccacc gcctcttccc cgatccctgg actccgactc cctggctttg1250 gcattcagtg agacgccctg cacacacaga aagccaaagc gatcggtgct 1300cccagatccc gcagcctctg gagagagctg acggcagatg aaatcaccag 1350 ggcggggcacccgcgagaac cctctgggac cttccgcggc cctctctgca 1400 cacatcctca agtgaccccgcacggcgaga cgcgggtggc ggcagggcgt 1450 cccagggtgc ggcaccgcgg ctccagtccttggaaataat taggcaaatt 1500 ctaaaggtct caaaaggagc aaagtaaacc gtggaggacaaagaaaaggg 1550 ttgttatttt tgtctttcca gccagcctgc tggctcccaa gagagaggcc1600 ttttcagttg agactctgct taagagaaga tccaaagtta aagctctggg 1650gtcaggggag gggccggggg caggaaacta cctctggctt aattctttta 1700 agccacgtaggaactttctt gagggatagg tggaccctga catccctgtg 1750 gccttgccca agggctctgctggtctttct gagtcacagc tgcgaggtga 1800 tgggggctgg ggccccaggc gtcagcctcccagagggaca gctgagcccc 1850 ctgccttggc tccaggttgg tagaagcagc cgaagggctcctgacagtgg 1900 ccagggaccc ctgggtcccc caggcctgca gatgtttcta tgaggggcag1950 agctccttgg tacatccatg tgtggctctg ctccacccct gtgccacccc 2000agagccctgg ggggtggtct ccatgcctgc caccctggca tcggctttct 2050 gtgccgcctcccacacaaat cagccccaga aggccccggg gccttggctt 2100 ctgtttttta taaaacacctcaagcagcac tgcagtctcc catctcctcg 2150 tgggctaagc atcaccgctt ccacgtgtgttgtgttggtt ggcagcaagg 2200 ctgatccaga ccccttctgc ccccactgcc ctcatccaggcctctgacca 2250 gtagcctgag aggggctttt tctaggcttc agagcagggg agagctggaa2300 ggggctagaa agctcccgct tgtctgtttc tcaggctcct gtgagcctca 2350gtcctgagac cagagtcaag aggaagtaca cgtcccaatc acccgtgtca 2400 ggattcactctcaggagctg ggtggcagga gaggcaatag cccctgtggc 2450 aattgcagga ccagctggagcagggttgcg gtgtctccac ggtgctctcg 2500 ccctgcccat ggccacccca gactctgatctccaggaacc ccatagcccc 2550 tctccacctc accccatgtt gatgcccagg gtcactcttgctacccgctg 2600 ggcccccaaa cccccgctgc ctctcttcct tccccccatc ccccacctgg2650 ttttgactaa tcctgcttcc ctctctgggc ctggctgccg ggatctgggg 2700tccctaagtc cctctcttta aagaacttct gcgggtcaga ctctgaagcc 2750 gagttgctgtgggcgtgccc ggaagcagag cgccacactc gctgcttaag 2800 ctcccccagc tctttccagaaaacattaaa ctcagaattg tgttttcaa 2849 73 281 PRT Homo Sapien 73 Met GlySer Arg Gly Gln Gly Leu Leu Leu Ala Tyr Cys Leu Leu 1 5 10 15 Leu AlaPhe Ala Ser Gly Leu Val Leu Ser Arg Val Pro His Val 20 25 30 Gln Gly GluGln Gln Glu Trp Glu Gly Thr Glu Glu Leu Pro Ser 35 40 45 Pro Pro Asp HisAla Glu Arg Ala Glu Glu Gln His Glu Lys Tyr 50 55 60 Arg Pro Ser Gln AspGln Gly Leu Pro Ala Ser Arg Cys Leu Arg 65 70 75 Cys Cys Asp Pro Gly ThrSer Met Tyr Pro Ala Thr Ala Val Pro 80 85 90 Gln Ile Asn Ile Thr Ile LeuLys Gly Glu Lys Gly Asp Arg Gly 95 100 105 Asp Arg Gly Leu Gln Gly LysTyr Gly Lys Thr Gly Ser Ala Gly 110 115 120 Ala Arg Gly His Thr Gly ProLys Gly Gln Lys Gly Ser Met Gly 125 130 135 Ala Pro Gly Glu Arg Cys LysSer His Tyr Ala Ala Phe Ser Val 140 145 150 Gly Arg Lys Lys Pro Met HisSer Asn His Tyr Tyr Gln Thr Val 155 160 165 Ile Phe Asp Thr Glu Phe ValAsn Leu Tyr Asp His Phe Asn Met 170 175 180 Phe Thr Gly Lys Phe Tyr CysTyr Val Pro Gly Leu Tyr Phe Phe 185 190 195 Ser Leu Asn Val His Thr TrpAsn Gln Lys Glu Thr Tyr Leu His 200 205 210 Ile Met Lys Asn Glu Glu GluVal Val Ile Leu Phe Ala Gln Val 215 220 225 Gly Asp Arg Ser Ile Met GlnSer Gln Ser Leu Met Leu Glu Leu 230 235 240 Arg Glu Gln Asp Gln Val TrpVal Arg Leu Tyr Lys Gly Glu Arg 245 250 255 Glu Asn Ala Ile Phe Ser GluGlu Leu Asp Thr Tyr Ile Thr Phe 260 265 270 Ser Gly Tyr Leu Val Lys HisAla Thr Glu Pro 275 280 74 24 DNA Artificial Sequence Syntheticoligonucleotide probe 74 tacaggccca gtcaggacca gggg 24 75 24 DNAArtificial Sequence Synthetic oligonucleotide probe 75 ctgaagaagtagaggccggg cacg 24 76 45 DNA Artificial Sequence Syntheticoligonucleotide probe 76 cccggtgctt gcgctgctgt gaccccggta cctccatgtacccgg 45 77 1042 DNA Homo Sapien 77 gaattcggca cgagggaaga agagaaagaaaatctccggg gctgctggga 50 gcatataaag aagccctgtg gccttgctgg ttttaccatccagaccagag 100 tcaggccaca gacggacatg gctgctcaag gctggtccat gctcctgctg150 gctgtcctta acctaggcat cttcgtccgt ccctgtgaca ctcaagagct 200acgatgtctg tgtattcagg aacactctga attcattcct ctcaaactca 250 ttaaaaatataatggtgata ttcgagacca tttactgcaa cagaaaggaa 300 gtgatagcag tcccaaaaaatgggagtatg atttgtttgg atcctgatgc 350 tccatgggtg aaggctactg ttggcccaattactaacagg ttcctacctg 400 aggacctcaa acaaaaggaa tttccaccgg caatgaagcttctgtatagt 450 gttgagcatg aaaagcctct atatctttca tttgggagac ctgagaacaa500 gagaatattt ccctttccaa ttcgggagac ctctagacac tttgctgatt 550tagctcacaa cagtgatagg aattttctac gggactccag tgaagtcagc 600 ttgacaggcagtgatgccta aaagccactc atgaggcaaa gagtttcaag 650 gaagctctcc tcctggagttttggcgttct cattcttata ctctattccc 700 gcgttagtct ggtgtatgga tctatgagctctcttttaat attttattat 750 aaatgtttta tttacttaac ttcctagtga atgttcacaggtgactgctc 800 ccccatcccc atttcttgat attacatata atggcatcat ataccccttt850 attgactgac aaactactca gattgcttaa cattttgtgc ttcaaagtct 900tatcccactc cactatgggc tgttacagag tgcatctcgg tgtagagcaa 950 ggctccttgtcttcagtgcc ccagggtgaa atacttcttt gaaaaatttt 1000 cattcatcag aaaatctgaaataaaaatat gtcttaattg ag 1042 78 167 PRT Homo Sapien 78 Met Ala Ala GlnGly Trp Ser Met Leu Leu Leu Ala Val Leu Asn 1 5 10 15 Leu Gly Ile PheVal Arg Pro Cys Asp Thr Gln Glu Leu Arg Cys 20 25 30 Leu Cys Ile Gln GluHis Ser Glu Phe Ile Pro Leu Lys Leu Ile 35 40 45 Lys Asn Ile Met Val IlePhe Glu Thr Ile Tyr Cys Asn Arg Lys 50 55 60 Glu Val Ile Ala Val Pro LysAsn Gly Ser Met Ile Cys Leu Asp 65 70 75 Pro Asp Ala Pro Trp Val Lys AlaThr Val Gly Pro Ile Thr Asn 80 85 90 Arg Phe Leu Pro Glu Asp Leu Lys GlnLys Glu Phe Pro Pro Ala 95 100 105 Met Lys Leu Leu Tyr Ser Val Glu HisGlu Lys Pro Leu Tyr Leu 110 115 120 Ser Phe Gly Arg Pro Glu Asn Lys ArgIle Phe Pro Phe Pro Ile 125 130 135 Arg Glu Thr Ser Arg His Phe Ala AspLeu Ala His Asn Ser Asp 140 145 150 Arg Asn Phe Leu Arg Asp Ser Ser GluVal Ser Leu Thr Gly Ser 155 160 165 Asp Ala 79 798 DNA Homo Sapienunsure 794 unknown base 79 cagacatggc tcagtcactg gctctgagcc tccttatcctggttctggcc 50 tttggcatcc ccaggaccca aggcagtgat ggaggggctc aggactgttg 100cctcaagtac agccaaagga agattcccgc caaggttgtc cgcagctacc 150 ggaagcaggaaccaagctta ggctgctcca tcccagctat cctgttcttg 200 ccccgcaagc gctctcaggcagagctatgt gcagacccaa aggagctctg 250 ggtgcagcag ctgatgcagc atctggacaagacaccatcc ccacagaaac 300 cagcccaggg ctgcaggaag gacagggggg cctccaagactggcaagaaa 350 ggaaagggct ccaaaggctg caagaggact gagcggtcac agacccctaa400 agggccatag cccagtgagc agcctggagc cctggagacc ccaccagcct 450caccagcgct tgaagcctga acccaagatg caagaaggag gctatgctca 500 ggggccctggagcagccacc ccatgctggc cttgccacac tctttctcct 550 gctttaacca ccccatctgcattcccagct ctaccctgca tggctgagct 600 gcccacagca ggccaggtcc agagagaccgaggagggaga gtctcccagg 650 gagcatgaga ggaggcagca ggactgtccc cttgaaggagaatcatcagg 700 accctggacc tgatacggct ccccagtaca ccccacctct tccttgtaaa750 tatgatttat acctaactga ataaaaagct gttctgtctt cccnccca 798 80 134 PRTHomo Sapien 80 Met Ala Gln Ser Leu Ala Leu Ser Leu Leu Ile Leu Val LeuAla 1 5 10 15 Phe Gly Ile Pro Arg Thr Gln Gly Ser Asp Gly Gly Ala GlnAsp 20 25 30 Cys Cys Leu Lys Tyr Ser Gln Arg Lys Ile Pro Ala Lys Val Val35 40 45 Arg Ser Tyr Arg Lys Gln Glu Pro Ser Leu Gly Cys Ser Ile Pro 5055 60 Ala Ile Leu Phe Leu Pro Arg Lys Arg Ser Gln Ala Glu Leu Cys 65 7075 Ala Asp Pro Lys Glu Leu Trp Val Gln Gln Leu Met Gln His Leu 80 85 90Asp Lys Thr Pro Ser Pro Gln Lys Pro Ala Gln Gly Cys Arg Lys 95 100 105Asp Arg Gly Ala Ser Lys Thr Gly Lys Lys Gly Lys Gly Ser Lys 110 115 120Gly Cys Lys Arg Thr Glu Arg Ser Gln Thr Pro Lys Gly Pro 125 130 81 20DNA Artificial Sequence Synthetic oligonucleotide probe 81 agacatggctcagtcactgg 20 82 19 DNA Artificial Sequence Synthetic oligonucleotideprobe 82 gacccctaaa gggccatag 19 83 924 DNA Homo Sapien 83 aaggagcagcccgcaagcac caagtgagag gcatgaagtt acagtgtgtt 50 tccctttggc tcctgggtacaatactgata ttgtgctcag tagacaacca 100 cggtctcagg agatgtctga tttccacagacatgcaccat atagaagaga 150 gtttccaaga aatcaaaaga gccatccaag ctaaggacaccttcccaaat 200 gtcactatcc tgtccacatt ggagactctg cagatcatta agcccttaga250 tgtgtgctgc gtgaccaaga acctcctggc gttctacgtg gacagggtgt 300tcaaggatca tcaggagcca aaccccaaaa tcttgagaaa aatcagcagc 350 attgccaactctttcctcta catgcagaaa actctgcggc aatgtcagga 400 acagaggcag tgtcactgcaggcaggaagc caccaatgcc accagagtca 450 tccatgacaa ctatgatcag ctggaggtccacgctgctgc cattaaatcc 500 ctgggagagc tcgacgtctt tctagcctgg attaataagaatcatgaagt 550 aatgttctca gcttgatgac aaggaacctg tatagtgatc cagggatgaa600 caccccctgt gcggtttact gtgggagaca gcccaccttg aaggggaagg 650agatggggaa ggccccttgc agctgaaagt cccactggct ggcctcaggc 700 tgtcttattccgcttgaaaa taggcaaaaa gtctactgtg gtatttgtaa 750 taaactctat ctgctgaaagggcctgcagg ccatcctggg agtaaagggc 800 tgccttccca tctaatttat tgtaaagtcatatagtccat gtctgtgatg 850 tgagccaagt gatatcctgt agtacacatt gtactgagtggtttttctga 900 ataaattcca tattttacct atga 924 84 177 PRT Homo Sapien 84Met Lys Leu Gln Cys Val Ser Leu Trp Leu Leu Gly Thr Ile Leu 1 5 10 15Ile Leu Cys Ser Val Asp Asn His Gly Leu Arg Arg Cys Leu Ile 20 25 30 SerThr Asp Met His His Ile Glu Glu Ser Phe Gln Glu Ile Lys 35 40 45 Arg AlaIle Gln Ala Lys Asp Thr Phe Pro Asn Val Thr Ile Leu 50 55 60 Ser Thr LeuGlu Thr Leu Gln Ile Ile Lys Pro Leu Asp Val Cys 65 70 75 Cys Val Thr LysAsn Leu Leu Ala Phe Tyr Val Asp Arg Val Phe 80 85 90 Lys Asp His Gln GluPro Asn Pro Lys Ile Leu Arg Lys Ile Ser 95 100 105 Ser Ile Ala Asn SerPhe Leu Tyr Met Gln Lys Thr Leu Arg Gln 110 115 120 Cys Gln Glu Gln ArgGln Cys His Cys Arg Gln Glu Ala Thr Asn 125 130 135 Ala Thr Arg Val IleHis Asp Asn Tyr Asp Gln Leu Glu Val His 140 145 150 Ala Ala Ala Ile LysSer Leu Gly Glu Leu Asp Val Phe Leu Ala 155 160 165 Trp Ile Asn Lys AsnHis Glu Val Met Phe Ser Ala 170 175 85 2137 DNA Homo Sapien 85gctcccagcc aagaacctcg gggccgctgc gcggtgggga ggagttcccc 50 gaaacccggccgctaagcga ggcctcctcc tcccgcagat ccgaacggcc 100 tgggcggggt caccccggctgggacaagaa gccgccgcct gcctgcccgg 150 gcccggggag ggggctgggg ctggggccggaggcggggtg tgagtgggtg 200 tgtgcggggg gcggaggctt gatgcaatcc cgataagaaatgctcgggtg 250 tcttgggcac ctacccgtgg ggcccgtaag gcgctactat ataaggctgc300 cggcccggag ccgccgcgcc gtcagagcag gagcgctgcg tccaggatct 350agggccacga ccatcccaac ccggcactca cagccccgca gcgcatcccg 400 gtcgccgcccagcctcccgc acccccatcg ccggagctgc gccgagagcc 450 ccagggaggt gccatgcggagcgggtgtgt ggtggtccac gtatggatcc 500 tggccggcct ctggctggcc gtggccgggcgccccctcgc cttctcggac 550 gcggggcccc acgtgcacta cggctggggc gaccccatccgcctgcggca 600 cctgtacacc tccggccccc acgggctctc cagctgcttc ctgcgcatcc650 gtgccgacgg cgtcgtggac tgcgcgcggg gccagagcgc gcacagtttg 700ctggagatca aggcagtcgc tctgcggacc gtggccatca agggcgtgca 750 cagcgtgcggtacctctgca tgggcgccga cggcaagatg caggggctgc 800 ttcagtactc ggaggaagactgtgctttcg aggaggagat ccgcccagat 850 ggctacaatg tgtaccgatc cgagaagcaccgcctcccgg tctccctgag 900 cagtgccaaa cagcggcagc tgtacaagaa cagaggctttcttccactct 950 ctcatttcct gcccatgctg cccatggtcc cagaggagcc tgaggacctc1000 aggggccact tggaatctga catgttctct tcgcccctgg agaccgacag 1050catggaccca tttgggcttg tcaccggact ggaggccgtg aggagtccca 1100 gctttgagaagtaactgaga ccatgcccgg gcctcttcac tgctgccagg 1150 ggctgtggta cctgcagcgtgggggacgtg cttctacaag aacagtcctg 1200 agtccacgtt ctgtttagct ttaggaagaaacatctagaa gttgtacata 1250 ttcagagttt tccattggca gtgccagttt ctagccaatagacttgtctg 1300 atcataacat tgtaagcctg tagcttgccc agctgctgcc tgggccccca1350 ttctgctccc tcgaggttgc tggacaagct gctgcactgt ctcagttctg 1400cttgaatacc tccatcgatg gggaactcac ttcctttgga aaaattctta 1450 tgtcaagctgaaattctcta attttttctc atcacttccc caggagcagc 1500 cagaagacag gcagtagttttaatttcagg aacaggtgat ccactctgta 1550 aaacagcagg taaatttcac tcaaccccatgtgggaattg atctatatct 1600 ctacttccag ggaccatttg cccttcccaa atccctccaggccagaactg 1650 actggagcag gcatggccca ccaggcttca ggagtagggg aagcctggag1700 ccccactcca gccctgggac aacttgagaa ttccccctga ggccagttct 1750gtcatggatg ctgtcctgag aataacttgc tgtcccggtg tcacctgctt 1800 ccatctcccagcccaccagc cctctgccca cctcacatgc ctccccatgg 1850 attggggcct cccaggccccccaccttatg tcaacctgca cttcttgttc 1900 aaaaatcagg aaaagaaaag atttgaagaccccaagtctt gtcaataact 1950 tgctgtgtgg aagcagcggg ggaagaccta gaaccctttccccagcactt 2000 ggttttccaa catgatattt atgagtaatt tattttgata tgtacatctc2050 ttattttctt acattattta tgcccccaaa ttatatttat gtatgtaagt 2100gaggtttgtt ttgtatatta aaatggagtt tgtttgt 2137 86 216 PRT Homo Sapien 86Met Arg Ser Gly Cys Val Val Val His Val Trp Ile Leu Ala Gly 1 5 10 15Leu Trp Leu Ala Val Ala Gly Arg Pro Leu Ala Phe Ser Asp Ala 20 25 30 GlyPro His Val His Tyr Gly Trp Gly Asp Pro Ile Arg Leu Arg 35 40 45 His LeuTyr Thr Ser Gly Pro His Gly Leu Ser Ser Cys Phe Leu 50 55 60 Arg Ile ArgAla Asp Gly Val Val Asp Cys Ala Arg Gly Gln Ser 65 70 75 Ala His Ser LeuLeu Glu Ile Lys Ala Val Ala Leu Arg Thr Val 80 85 90 Ala Ile Lys Gly ValHis Ser Val Arg Tyr Leu Cys Met Gly Ala 95 100 105 Asp Gly Lys Met GlnGly Leu Leu Gln Tyr Ser Glu Glu Asp Cys 110 115 120 Ala Phe Glu Glu GluIle Arg Pro Asp Gly Tyr Asn Val Tyr Arg 125 130 135 Ser Glu Lys His ArgLeu Pro Val Ser Leu Ser Ser Ala Lys Gln 140 145 150 Arg Gln Leu Tyr LysAsn Arg Gly Phe Leu Pro Leu Ser His Phe 155 160 165 Leu Pro Met Leu ProMet Val Pro Glu Glu Pro Glu Asp Leu Arg 170 175 180 Gly His Leu Glu SerAsp Met Phe Ser Ser Pro Leu Glu Thr Asp 185 190 195 Ser Met Asp Pro PheGly Leu Val Thr Gly Leu Glu Ala Val Arg 200 205 210 Ser Pro Ser Phe GluLys 215 87 26 DNA Artificial Sequence Synthetic oligonucleotide probe 87atccgcccag atggctacaa tgtgta 26 88 42 DNA Artificial Sequence Syntheticoligonucleotide probe 88 gcctcccggt ctccctgagc agtgccaaac agcggcagtg ta42 89 22 DNA Artificial Sequence Synthetic oligonucleotide probe 89ccagtccggt gacaagccca aa 22 90 1857 DNA Homo Sapien 90 gtctgttcccaggagtcctt cggcggctgt tgtgtcagtg gcctgatcgc 50 gatggggaca aaggcgcaagtcgagaggaa actgttgtgc ctcttcatat 100 tggcgatcct gttgtgctcc ctggcattgggcagtgttac agtgcactct 150 tctgaacctg aagtcagaat tcctgagaat aatcctgtgaagttgtcctg 200 tgcctactcg ggcttttctt ctccccgtgt ggagtggaag tttgaccaag250 gagacaccac cagactcgtt tgctataata acaagatcac agcttcctat 300gaggaccggg tgaccttctt gccaactggt atcaccttca agtccgtgac 350 acgggaagacactgggacat acacttgtat ggtctctgag gaaggcggca 400 acagctatgg ggaggtcaaggtcaagctca tcgtgcttgt gcctccatcc 450 aagcctacag ttaacatccc ctcctctgccaccattggga accgggcagt 500 gctgacatgc tcagaacaag atggttcccc accttctgaatacacctggt 550 tcaaagatgg gatagtgatg cctacgaatc ccaaaagcac ccgtgccttc600 agcaactctt cctatgtcct gaatcccaca acaggagagc tggtctttga 650tcccctgtca gcctctgata ctggagaata cagctgtgag gcacggaatg 700 ggtatgggacacccatgact tcaaatgctg tgcgcatgga agctgtggag 750 cggaatgtgg gggtcatcgtggcagccgtc cttgtaaccc tgattctcct 800 gggaatcttg gtttttggca tctggtttgcctatagccga ggccactttg 850 acagaacaaa gaaagggact tcgagtaaga aggtgatttacagccagcct 900 agtgcccgaa gtgaaggaga attcaaacag acctcgtcat tcctggtgtg950 agcctggtcg gctcaccgcc tatcatctgc atttgcctta ctcaggtgct 1000accggactct ggcccctgat gtctgtagtt tcacaggatg ccttatttgt 1050 cttctacaccccacagggcc ccctacttct tcggatgtgt ttttaataat 1100 gtcagctatg tgccccatcctccttcatgc cctccctccc tttcctacca 1150 ctgctgagtg gcctggaact tgtttaaagtgtttattccc catttctttg 1200 agggatcagg aaggaatcct gggtatgcca ttgacttcccttctaagtag 1250 acagcaaaaa tggcgggggt cgcaggaatc tgcactcaac tgcccacctg1300 gctggcaggg atctttgaat aggtatcttg agcttggttc tgggctcttt 1350ccttgtgtac tgacgaccag ggccagctgt tctagagcgg gaattagagg 1400 ctagagcggctgaaatggtt gtttggtgat gacactgggg tccttccatc 1450 tctggggccc actctcttctgtcttcccat gggaagtgcc actgggatcc 1500 ctctgccctg tcctcctgaa tacaagctgactgacattga ctgtgtctgt 1550 ggaaaatggg agctcttgtt gtggagagca tagtaaattttcagagaact 1600 tgaagccaaa aggatttaaa accgctgctc taaagaaaag aaaactggag1650 gctgggcgca gtggctcacg cctgtaatcc cagaggctga ggcaggcgga 1700tcacctgagg tcgggagttc gggatcagcc tgaccaacat ggagaaaccc 1750 tactggaaatacaaagttag ccaggcatgg tggtgcatgc ctgtagtccc 1800 agctgctcag gagcctggcaacaagagcaa aactccagct caaaaaaaaa 1850 aaaaaaa 1857 91 299 PRT HomoSapien 91 Met Gly Thr Lys Ala Gln Val Glu Arg Lys Leu Leu Cys Leu Phe 15 10 15 Ile Leu Ala Ile Leu Leu Cys Ser Leu Ala Leu Gly Ser Val Thr 2025 30 Val His Ser Ser Glu Pro Glu Val Arg Ile Pro Glu Asn Asn Pro 35 4045 Val Lys Leu Ser Cys Ala Tyr Ser Gly Phe Ser Ser Pro Arg Val 50 55 60Glu Trp Lys Phe Asp Gln Gly Asp Thr Thr Arg Leu Val Cys Tyr 65 70 75 AsnAsn Lys Ile Thr Ala Ser Tyr Glu Asp Arg Val Thr Phe Leu 80 85 90 Pro ThrGly Ile Thr Phe Lys Ser Val Thr Arg Glu Asp Thr Gly 95 100 105 Thr TyrThr Cys Met Val Ser Glu Glu Gly Gly Asn Ser Tyr Gly 110 115 120 Glu ValLys Val Lys Leu Ile Val Leu Val Pro Pro Ser Lys Pro 125 130 135 Thr ValAsn Ile Pro Ser Ser Ala Thr Ile Gly Asn Arg Ala Val 140 145 150 Leu ThrCys Ser Glu Gln Asp Gly Ser Pro Pro Ser Glu Tyr Thr 155 160 165 Trp PheLys Asp Gly Ile Val Met Pro Thr Asn Pro Lys Ser Thr 170 175 180 Arg AlaPhe Ser Asn Ser Ser Tyr Val Leu Asn Pro Thr Thr Gly 185 190 195 Glu LeuVal Phe Asp Pro Leu Ser Ala Ser Asp Thr Gly Glu Tyr 200 205 210 Ser CysGlu Ala Arg Asn Gly Tyr Gly Thr Pro Met Thr Ser Asn 215 220 225 Ala ValArg Met Glu Ala Val Glu Arg Asn Val Gly Val Ile Val 230 235 240 Ala AlaVal Leu Val Thr Leu Ile Leu Leu Gly Ile Leu Val Phe 245 250 255 Gly IleTrp Phe Ala Tyr Ser Arg Gly His Phe Asp Arg Thr Lys 260 265 270 Lys GlyThr Ser Ser Lys Lys Val Ile Tyr Ser Gln Pro Ser Ala 275 280 285 Arg SerGlu Gly Glu Phe Lys Gln Thr Ser Ser Phe Leu Val 290 295 92 24 DNAArtificial Sequence Synthetic oligonucleotide probe 92 tcgcggagctgtgttctgtt tccc 24 93 50 DNA Artificial Sequence Syntheticoligonucleotide probe 93 tgatcgcgat ggggacaaag gcgcaagctc gagaggaaactgttgtgcct 50 94 20 DNA Artificial Sequence Synthetic oligonucleotideprobe 94 acacctggtt caaagatggg 20 95 24 DNA Artificial SequenceSynthetic oligonucleotide probe 95 taggaagagt tgctgaaggc acgg 24 96 20DNA Artificial Sequence Synthetic oligonucleotide probe 96 ttgccttactcaggtgctac 20 97 20 DNA Artificial Sequence Synthetic oligonucleotideprobe 97 actcagcagt ggtaggaaag 20 98 1200 DNA Homo Sapien 98 cccacgcgtccgaacctctc cagcgatggg agccgcccgc ctgctgccca 50 acctcactct gtgcttacagctgctgattc tctgctgtca aactcagtac 100 gtgagggacc agggcgccat gaccgaccagctgagcaggc ggcagatccg 150 cgagtaccaa ctctacagca ggaccagtgg caagcacgtgcaggtcaccg 200 ggcgtcgcat ctccgccacc gccgaggacg gcaacaagtt tgccaagctc250 atagtggaga cggacacgtt tggcagccgg gttcgcatca aaggggctga 300gagtgagaag tacatctgta tgaacaagag gggcaagctc atcgggaagc 350 ccagcgggaagagcaaagac tgcgtgttca cggagatcgt gctggagaac 400 aactatacgg ccttccagaacgcccggcac gagggctggt tcatggcctt 450 cacgcggcag gggcggcccc gccaggcttcccgcagccgc cagaaccagc 500 gcgaggccca cttcatcaag cgcctctacc aaggccagctgcccttcccc 550 aaccacgccg agaagcagaa gcagttcgag tttgtgggct ccgcccccac600 ccgccggacc aagcgcacac ggcggcccca gcccctcacg tagtctggga 650ggcagggggc agcagcccct gggccgcctc cccacccctt tcccttctta 700 atccaaggactgggctgggg tggcgggagg ggagccagat ccccgaggga 750 ggaccctgag ggccgcgaagcatccgagcc cccagctggg aaggggcagg 800 ccggtgcccc aggggcggct ggcacagtgcccccttcccg gacgggtggc 850 aggccctgga gaggaactga gtgtcaccct gatctcaggccaccagcctc 900 tgccggcctc ccagccgggc tcctgaagcc cgctgaaagg tcagcgactg950 aaggccttgc agacaaccgt ctggaggtgg ctgtcctcaa aatctgcttc 1000tcggatctcc ctcagtctgc ccccagcccc caaactcctc ctggctagac 1050 tgtaggaagggacttttgtt tgtttgtttg tttcaggaaa aaagaaaggg 1100 agagagagga aaatagagggttgtccactc ctcacattcc acgacccagg 1150 cctgcacccc acccccaact cccagccccggaataaaacc attttcctgc 1200 99 205 PRT Homo Sapien 99 Met Gly Ala Ala ArgLeu Leu Pro Asn Leu Thr Leu Cys Leu Gln 1 5 10 15 Leu Leu Ile Leu CysCys Gln Thr Gln Tyr Val Arg Asp Gln Gly 20 25 30 Ala Met Thr Asp Gln LeuSer Arg Arg Gln Ile Arg Glu Tyr Gln 35 40 45 Leu Tyr Ser Arg Thr Ser GlyLys His Val Gln Val Thr Gly Arg 50 55 60 Arg Ile Ser Ala Thr Ala Glu AspGly Asn Lys Phe Ala Lys Leu 65 70 75 Ile Val Glu Thr Asp Thr Phe Gly SerArg Val Arg Ile Lys Gly 80 85 90 Ala Glu Ser Glu Lys Tyr Ile Cys Met AsnLys Arg Gly Lys Leu 95 100 105 Ile Gly Lys Pro Ser Gly Lys Ser Lys AspCys Val Phe Thr Glu 110 115 120 Ile Val Leu Glu Asn Asn Tyr Thr Ala PheGln Asn Ala Arg His 125 130 135 Glu Gly Trp Phe Met Ala Phe Thr Arg GlnGly Arg Pro Arg Gln 140 145 150 Ala Ser Arg Ser Arg Gln Asn Gln Arg GluAla His Phe Ile Lys 155 160 165 Arg Leu Tyr Gln Gly Gln Leu Pro Phe ProAsn His Ala Glu Lys 170 175 180 Gln Lys Gln Phe Glu Phe Val Gly Ser AlaPro Thr Arg Arg Thr 185 190 195 Lys Arg Thr Arg Arg Pro Gln Pro Leu Thr200 205 100 28 DNA Artificial Sequence Synthetic oligonucleotide probe100 cagtacgtga gggaccaggg cgccatga 28 101 24 DNA Artificial SequenceSynthetic oligonucleotide probe 101 ccggtgacct gcacgtgctt gcca 24 102 41DNA Artificial Sequence Synthetic oligonucleotide probe 102 gcggatctgccgcctgctca nctggtcggt catggcgccc t 41 103 1679 DNA Homo Sapien 103gttgtgtcct tcagcaaaac agtggattta aatctccttg cacaagcttg 50 agagcaacacaatctatcag gaaagaaaga aagaaaaaaa ccgaacctga 100 caaaaaagaa gaaaaagaagaagaaaaaaa atcatgaaaa ccatccagcc 150 aaaaatgcac aattctatct cttgggcaatcttcacgggg ctggctgctc 200 tgtgtctctt ccaaggagtg cccgtgcgca gcggagatgccaccttcccc 250 aaagctatgg acaacgtgac ggtccggcag ggggagagcg ccaccctcag300 gtgcactatt gacaaccggg tcacccgggt ggcctggcta aaccgcagca 350ccatcctcta tgctgggaat gacaagtggt gcctggatcc tcgcgtggtc 400 cttctgagcaacacccaaac gcagtacagc atcgagatcc agaacgtgga 450 tgtgtatgac gagggcccttacacctgctc ggtgcagaca gacaaccacc 500 caaagacctc tagggtccac ctcattgtgcaagtatctcc caaaattgta 550 gagatttctt cagatatctc cattaatgaa gggaacaatattagcctcac 600 ctgcatagca actggtagac cagagcctac ggttacttgg agacacatct650 ctcccaaagc ggttggcttt gtgagtgaag acgaatactt ggaaattcag 700ggcatcaccc gggagcagtc aggggactac gagtgcagtg cctccaatga 750 cgtggccgcgcccgtggtac ggagagtaaa ggtcaccgtg aactatccac 800 catacatttc agaagccaagggtacaggtg tccccgtggg acaaaagggg 850 acactgcagt gtgaagcctc agcagtcccctcagcagaat tccagtggta 900 caaggatgac aaaagactga ttgaaggaaa gaaaggggtgaaagtggaaa 950 acagaccttt cctctcaaaa ctcatcttct tcaatgtctc tgaacatgac1000 tatgggaact acacttgcgt ggcctccaac aagctgggcc acaccaatgc 1050cagcatcatg ctatttggtc caggcgccgt cagcgaggtg agcaacggca 1100 cgtcgaggagggcaggctgc gtctggctgc tgcctcttct ggtcttgcac 1150 ctgcttctca aattttgatgtgagtgccac ttccccaccc gggaaaggct 1200 gccgccacca ccaccaccaa cacaacagcaatggcaacac cgacagcaac 1250 caatcagata tatacaaatg aaattagaag aaacacagcctcatgggaca 1300 gaaatttgag ggaggggaac aaagaatact ttggggggaa aagagtttta1350 aaaaagaaat tgaaaattgc cttgcagata tttaggtaca atggagtttt 1400cttttcccaa acgggaagaa cacagcacac ccggcttgga cccactgcaa 1450 gctgcatcgtgcaacctctt tggtgccagt gtgggcaagg gctcagcctc 1500 tctgcccaca gagtgcccccacgtggaaca ttctggagct ggccatccca 1550 aattcaatca gtccatagag acgaacagaatgagaccttc cggcccaagc 1600 gtggcgctgc gggcactttg gtagactgtg ccaccacggcgtgtgttgtg 1650 aaacgtgaaa taaaaagagc aaaaaaaaa 1679 104 344 PRT HomoSapien 104 Met Lys Thr Ile Gln Pro Lys Met His Asn Ser Ile Ser Trp Ala 15 10 15 Ile Phe Thr Gly Leu Ala Ala Leu Cys Leu Phe Gln Gly Val Pro 2025 30 Val Arg Ser Gly Asp Ala Thr Phe Pro Lys Ala Met Asp Asn Val 35 4045 Thr Val Arg Gln Gly Glu Ser Ala Thr Leu Arg Cys Thr Ile Asp 50 55 60Asn Arg Val Thr Arg Val Ala Trp Leu Asn Arg Ser Thr Ile Leu 65 70 75 TyrAla Gly Asn Asp Lys Trp Cys Leu Asp Pro Arg Val Val Leu 80 85 90 Leu SerAsn Thr Gln Thr Gln Tyr Ser Ile Glu Ile Gln Asn Val 95 100 105 Asp ValTyr Asp Glu Gly Pro Tyr Thr Cys Ser Val Gln Thr Asp 110 115 120 Asn HisPro Lys Thr Ser Arg Val His Leu Ile Val Gln Val Ser 125 130 135 Pro LysIle Val Glu Ile Ser Ser Asp Ile Ser Ile Asn Glu Gly 140 145 150 Asn AsnIle Ser Leu Thr Cys Ile Ala Thr Gly Arg Pro Glu Pro 155 160 165 Thr ValThr Trp Arg His Ile Ser Pro Lys Ala Val Gly Phe Val 170 175 180 Ser GluAsp Glu Tyr Leu Glu Ile Gln Gly Ile Thr Arg Glu Gln 185 190 195 Ser GlyAsp Tyr Glu Cys Ser Ala Ser Asn Asp Val Ala Ala Pro 200 205 210 Val ValArg Arg Val Lys Val Thr Val Asn Tyr Pro Pro Tyr Ile 215 220 225 Ser GluAla Lys Gly Thr Gly Val Pro Val Gly Gln Lys Gly Thr 230 235 240 Leu GlnCys Glu Ala Ser Ala Val Pro Ser Ala Glu Phe Gln Trp 245 250 255 Tyr LysAsp Asp Lys Arg Leu Ile Glu Gly Lys Lys Gly Val Lys 260 265 270 Val GluAsn Arg Pro Phe Leu Ser Lys Leu Ile Phe Phe Asn Val 275 280 285 Ser GluHis Asp Tyr Gly Asn Tyr Thr Cys Val Ala Ser Asn Lys 290 295 300 Leu GlyHis Thr Asn Ala Ser Ile Met Leu Phe Gly Pro Gly Ala 305 310 315 Val SerGlu Val Ser Asn Gly Thr Ser Arg Arg Ala Gly Cys Val 320 325 330 Trp LeuLeu Pro Leu Leu Val Leu His Leu Leu Leu Lys Phe 335 340 105 1734 DNAHomo Sapien 105 gtggactctg agaagcccag gcagttgagg acaggagaga gaaggctgca50 gacccagagg gagggaggac agggagtcgg aaggaggagg acagaggagg 100 gcacagagacgcagagcaag ggcggcaagg aggagaccct ggtgggagga 150 agacactctg gagagagagggggctgggca gagatgaagt tccaggggcc 200 cctggcctgc ctcctgctgg ccctctgcctgggcagtggg gaggctggcc 250 ccctgcagag cggagaggaa agcactggga caaatattggggaggccctt 300 ggacatggcc tgggagacgc cctgagcgaa ggggtgggaa aggccattgg350 caaagaggcc ggaggggcag ctggctctaa agtcagtgag gcccttggcc 400aagggaccag agaagcagtt ggcactggag tcaggcaggt tccaggcttt 450 ggcgcagcagatgctttggg caacagggtc ggggaagcag cccatgctct 500 gggaaacact gggcacgagattggcagaca ggcagaagat gtcattcgac 550 acggagcaga tgctgtccgc ggctcctggcagggggtgcc tggccacagt 600 ggtgcttggg aaacttctgg aggccatggc atctttggctctcaaggtgg 650 ccttggaggc cagggccagg gcaatcctgg aggtctgggg actccgtggg700 tccacggata ccccggaaac tcagcaggca gctttggaat gaatcctcag 750ggagctccct ggggtcaagg aggcaatgga gggccaccaa actttgggac 800 caacactcagggagctgtgg cccagcctgg ctatggttca gtgagagcca 850 gcaaccagaa tgaagggtgcacgaatcccc caccatctgg ctcaggtgga 900 ggctccagca actctggggg aggcagcggctcacagtcgg gcagcagtgg 950 cagtggcagc aatggtgaca acaacaatgg cagcagcagtggtggcagca 1000 gcagtggcag cagcagtggc agcagcagtg gcggcagcag tggcggcagc1050 agtggtggca gcagtggcaa cagtggtggc agcagaggtg acagcggcag 1100tgagtcctcc tggggatcca gcaccggctc ctcctccggc aaccacggtg 1150 ggagcggcggaggaaatgga cataaacccg ggtgtgaaaa gccagggaat 1200 gaagcccgcg ggagcggggaatctgggatt cagggcttca gaggacaggg 1250 agtttccagc aacatgaggg aaataagcaaagagggcaat cgcctccttg 1300 gaggctctgg agacaattat cgggggcaag ggtcgagctggggcagtgga 1350 ggaggtgacg ctgttggtgg agtcaatact gtgaactctg agacgtctcc1400 tgggatgttt aactttgaca ctttctggaa gaattttaaa tccaagctgg 1450gtttcatcaa ctgggatgcc ataaacaagg accagagaag ctctcgcatc 1500 ccgtgacctccagacaagga gccaccagat tggatgggag cccccacact 1550 ccctccttaa aacaccaccctctcatcact aatctcagcc cttgcccttg 1600 aaataaacct tagctgcccc acaaaaaaaaaaaaaaaaaa aaaaaaaaaa 1650 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaaaaaaaaaaaa 1700 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaa 1734 106 440 PRTHomo Sapien 106 Met Lys Phe Gln Gly Pro Leu Ala Cys Leu Leu Leu Ala LeuCys 1 5 10 15 Leu Gly Ser Gly Glu Ala Gly Pro Leu Gln Ser Gly Glu GluSer 20 25 30 Thr Gly Thr Asn Ile Gly Glu Ala Leu Gly His Gly Leu Gly Asp35 40 45 Ala Leu Ser Glu Gly Val Gly Lys Ala Ile Gly Lys Glu Ala Gly 5055 60 Gly Ala Ala Gly Ser Lys Val Ser Glu Ala Leu Gly Gln Gly Thr 65 7075 Arg Glu Ala Val Gly Thr Gly Val Arg Gln Val Pro Gly Phe Gly 80 85 90Ala Ala Asp Ala Leu Gly Asn Arg Val Gly Glu Ala Ala His Ala 95 100 105Leu Gly Asn Thr Gly His Glu Ile Gly Arg Gln Ala Glu Asp Val 110 115 120Ile Arg His Gly Ala Asp Ala Val Arg Gly Ser Trp Gln Gly Val 125 130 135Pro Gly His Ser Gly Ala Trp Glu Thr Ser Gly Gly His Gly Ile 140 145 150Phe Gly Ser Gln Gly Gly Leu Gly Gly Gln Gly Gln Gly Asn Pro 155 160 165Gly Gly Leu Gly Thr Pro Trp Val His Gly Tyr Pro Gly Asn Ser 170 175 180Ala Gly Ser Phe Gly Met Asn Pro Gln Gly Ala Pro Trp Gly Gln 185 190 195Gly Gly Asn Gly Gly Pro Pro Asn Phe Gly Thr Asn Thr Gln Gly 200 205 210Ala Val Ala Gln Pro Gly Tyr Gly Ser Val Arg Ala Ser Asn Gln 215 220 225Asn Glu Gly Cys Thr Asn Pro Pro Pro Ser Gly Ser Gly Gly Gly 230 235 240Ser Ser Asn Ser Gly Gly Gly Ser Gly Ser Gln Ser Gly Ser Ser 245 250 255Gly Ser Gly Ser Asn Gly Asp Asn Asn Asn Gly Ser Ser Ser Gly 260 265 270Gly Ser Ser Ser Gly Ser Ser Ser Gly Ser Ser Ser Gly Gly Ser 275 280 285Ser Gly Gly Ser Ser Gly Gly Ser Ser Gly Asn Ser Gly Gly Ser 290 295 300Arg Gly Asp Ser Gly Ser Glu Ser Ser Trp Gly Ser Ser Thr Gly 305 310 315Ser Ser Ser Gly Asn His Gly Gly Ser Gly Gly Gly Asn Gly His 320 325 330Lys Pro Gly Cys Glu Lys Pro Gly Asn Glu Ala Arg Gly Ser Gly 335 340 345Glu Ser Gly Ile Gln Gly Phe Arg Gly Gln Gly Val Ser Ser Asn 350 355 360Met Arg Glu Ile Ser Lys Glu Gly Asn Arg Leu Leu Gly Gly Ser 365 370 375Gly Asp Asn Tyr Arg Gly Gln Gly Ser Ser Trp Gly Ser Gly Gly 380 385 390Gly Asp Ala Val Gly Gly Val Asn Thr Val Asn Ser Glu Thr Ser 395 400 405Pro Gly Met Phe Asn Phe Asp Thr Phe Trp Lys Asn Phe Lys Ser 410 415 420Lys Leu Gly Phe Ile Asn Trp Asp Ala Ile Asn Lys Asp Gln Arg 425 430 435Ser Ser Arg Ile Pro 440 107 918 DNA Homo Sapien 107 agccaggcagcacatcacag cgggaggagc tgtcccaggt ggcccagctc 50 agcaatggca atgggggtccccagagtcat tctgctctgc ctctttgggg 100 ctgcgctctg cctgacaggg tcccaagccctgcagtgcta cagctttgag 150 cacacctact ttggcccctt tgacctcagg gccatgaagctgcccagcat 200 ctcctgtcct catgagtgct ttgaggctat cctgtctctg gacaccgggt250 atcgcgcgcc ggtgaccctg gtgcggaagg gctgctggac cgggcctcct 300gcgggccaga cgcaatcgaa cccggacgcg ctgccgccag actactcggt 350 ggtgcgcggctgcacaactg acaaatgcaa cgcccacctc atgactcatg 400 acgccctccc caacctgagccaagcacccg acccgccgac gctcagcggc 450 gccgagtgct acgcctgtat cggggtccaccaggatgact gcgctatcgg 500 caggtcccga cgagtccagt gtcaccagga ccagaccgcctgcttccagg 550 gcagtggcag aatgacagtt ggcaatttct cagtccctgt gtacatcaga600 acctgccacc ggccctcctg caccaccgag ggcaccacca gcccctggac 650agccatcgac ctccagggct cctgctgtga ggggtacctc tgcaacagga 700 aatccatgacccagcccttc accagtgctt cagccaccac ccctccccga 750 gcactacagg tcctggccctgctcctccca gtcctcctgc tggtggggct 800 ctcagcatag accgcccctc caggatgctggggacagggc tcacacacct 850 cattcttgct gcttcagccc ctatcacata gctcactggaaaatgatgtt 900 aaagtaagaa ttgcaaaa 918 108 251 PRT Homo Sapien 108 MetAla Met Gly Val Pro Arg Val Ile Leu Leu Cys Leu Phe Gly 1 5 10 15 AlaAla Leu Cys Leu Thr Gly Ser Gln Ala Leu Gln Cys Tyr Ser 20 25 30 Phe GluHis Thr Tyr Phe Gly Pro Phe Asp Leu Arg Ala Met Lys 35 40 45 Leu Pro SerIle Ser Cys Pro His Glu Cys Phe Glu Ala Ile Leu 50 55 60 Ser Leu Asp ThrGly Tyr Arg Ala Pro Val Thr Leu Val Arg Lys 65 70 75 Gly Cys Trp Thr GlyPro Pro Ala Gly Gln Thr Gln Ser Asn Pro 80 85 90 Asp Ala Leu Pro Pro AspTyr Ser Val Val Arg Gly Cys Thr Thr 95 100 105 Asp Lys Cys Asn Ala HisLeu Met Thr His Asp Ala Leu Pro Asn 110 115 120 Leu Ser Gln Ala Pro AspPro Pro Thr Leu Ser Gly Ala Glu Cys 125 130 135 Tyr Ala Cys Ile Gly ValHis Gln Asp Asp Cys Ala Ile Gly Arg 140 145 150 Ser Arg Arg Val Gln CysHis Gln Asp Gln Thr Ala Cys Phe Gln 155 160 165 Gly Ser Gly Arg Met ThrVal Gly Asn Phe Ser Val Pro Val Tyr 170 175 180 Ile Arg Thr Cys His ArgPro Ser Cys Thr Thr Glu Gly Thr Thr 185 190 195 Ser Pro Trp Thr Ala IleAsp Leu Gln Gly Ser Cys Cys Glu Gly 200 205 210 Tyr Leu Cys Asn Arg LysSer Met Thr Gln Pro Phe Thr Ser Ala 215 220 225 Ser Ala Thr Thr Pro ProArg Ala Leu Gln Val Leu Ala Leu Leu 230 235 240 Leu Pro Val Leu Leu LeuVal Gly Leu Ser Ala 245 250 109 1813 DNA Homo Sapien 109 ggagccgccctgggtgtcag cggctcggct cccgcgcacg ctccggccgt 50 cgcgcagcct cggcacctgcaggtccgtgc gtcccgcggc tggcgcccct 100 gactccgtcc cggccaggga gggccatgatttccctcccg gggcccctgg 150 tgaccaactt gctgcggttt ttgttcctgg ggctgagtgccctcgcgccc 200 ccctcgcggg cccagctgca actgcacttg cccgccaacc ggttgcaggc250 ggtggaggga ggggaagtgg tgcttccagc gtggtacacc ttgcacgggg 300aggtgtcttc atcccagcca tgggaggtgc cctttgtgat gtggttcttc 350 aaacagaaagaaaaggagga tcaggtgttg tcctacatca atggggtcac 400 aacaagcaaa cctggagtatccttggtcta ctccatgccc tcccggaacc 450 tgtccctgcg gctggagggt ctccaggagaaagactctgg cccctacagc 500 tgctccgtga atgtgcaaga caaacaaggc aaatctaggggccacagcat 550 caaaacctta gaactcaatg tactggttcc tccagctcct ccatcctgcc600 gtctccaggg tgtgccccat gtgggggcaa acgtgaccct gagctgccag 650tctccaagga gtaagcccgc tgtccaatac cagtgggatc ggcagcttcc 700 atccttccagactttctttg caccagcatt agatgtcatc cgtgggtctt 750 taagcctcac caacctttcgtcttccatgg ctggagtcta tgtctgcaag 800 gcccacaatg aggtgggcac tgcccaatgtaatgtgacgc tggaagtgag 850 cacagggcct ggagctgcag tggttgctgg agctgttgtgggtaccctgg 900 ttggactggg gttgctggct gggctggtcc tcttgtacca ccgccggggc950 aaggccctgg aggagccagc caatgatatc aaggaggatg ccattgctcc 1000ccggaccctg ccctggccca agagctcaga cacaatctcc aagaatggga 1050 ccctttcctctgtcacctcc gcacgagccc tccggccacc ccatggccct 1100 cccaggcctg gtgcattgacccccacgccc agtctctcca gccaggccct 1150 gccctcacca agactgccca cgacagatggggcccaccct caaccaatat 1200 cccccatccc tggtggggtt tcttcctctg gcttgagccgcatgggtgct 1250 gtgcctgtga tggtgcctgc ccagagtcaa gctggctctc tggtatgatg1300 accccaccac tcattggcta aaggatttgg ggtctctcct tcctataagg 1350gtcacctcta gcacagaggc ctgagtcatg ggaaagagtc acactcctga 1400 cccttagtactctgccccca cctctcttta ctgtgggaaa accatctcag 1450 taagacctaa gtgtccaggagacagaagga gaagaggaag tggatctgga 1500 attgggagga gcctccaccc acccctgactcctccttatg aagccagctg 1550 ctgaaattag ctactcacca agagtgaggg gcagagacttccagtcactg 1600 agtctcccag gcccccttga tctgtacccc acccctatct aacaccaccc1650 ttggctccca ctccagctcc ctgtattgat ataacctgtc aggctggctt 1700ggttaggttt tactggggca gaggataggg aatctcttat taaaactaac 1750 atgaaatatgtgttgttttc atttgcaaat ttaaataaag atacataatg 1800 tttgtatgaa aaa 1813 110390 PRT Homo Sapien 110 Met Ile Ser Leu Pro Gly Pro Leu Val Thr Asn LeuLeu Arg Phe 1 5 10 15 Leu Phe Leu Gly Leu Ser Ala Leu Ala Pro Pro SerArg Ala Gln 20 25 30 Leu Gln Leu His Leu Pro Ala Asn Arg Leu Gln Ala ValGlu Gly 35 40 45 Gly Glu Val Val Leu Pro Ala Trp Tyr Thr Leu His Gly GluVal 50 55 60 Ser Ser Ser Gln Pro Trp Glu Val Pro Phe Val Met Trp Phe Phe65 70 75 Lys Gln Lys Glu Lys Glu Asp Gln Val Leu Ser Tyr Ile Asn Gly 8085 90 Val Thr Thr Ser Lys Pro Gly Val Ser Leu Val Tyr Ser Met Pro 95 100105 Ser Arg Asn Leu Ser Leu Arg Leu Glu Gly Leu Gln Glu Lys Asp 110 115120 Ser Gly Pro Tyr Ser Cys Ser Val Asn Val Gln Asp Lys Gln Gly 125 130135 Lys Ser Arg Gly His Ser Ile Lys Thr Leu Glu Leu Asn Val Leu 140 145150 Val Pro Pro Ala Pro Pro Ser Cys Arg Leu Gln Gly Val Pro His 155 160165 Val Gly Ala Asn Val Thr Leu Ser Cys Gln Ser Pro Arg Ser Lys 170 175180 Pro Ala Val Gln Tyr Gln Trp Asp Arg Gln Leu Pro Ser Phe Gln 185 190195 Thr Phe Phe Ala Pro Ala Leu Asp Val Ile Arg Gly Ser Leu Ser 200 205210 Leu Thr Asn Leu Ser Ser Ser Met Ala Gly Val Tyr Val Cys Lys 215 220225 Ala His Asn Glu Val Gly Thr Ala Gln Cys Asn Val Thr Leu Glu 230 235240 Val Ser Thr Gly Pro Gly Ala Ala Val Val Ala Gly Ala Val Val 245 250255 Gly Thr Leu Val Gly Leu Gly Leu Leu Ala Gly Leu Val Leu Leu 260 265270 Tyr His Arg Arg Gly Lys Ala Leu Glu Glu Pro Ala Asn Asp Ile 275 280285 Lys Glu Asp Ala Ile Ala Pro Arg Thr Leu Pro Trp Pro Lys Ser 290 295300 Ser Asp Thr Ile Ser Lys Asn Gly Thr Leu Ser Ser Val Thr Ser 305 310315 Ala Arg Ala Leu Arg Pro Pro His Gly Pro Pro Arg Pro Gly Ala 320 325330 Leu Thr Pro Thr Pro Ser Leu Ser Ser Gln Ala Leu Pro Ser Pro 335 340345 Arg Leu Pro Thr Thr Asp Gly Ala His Pro Gln Pro Ile Ser Pro 350 355360 Ile Pro Gly Gly Val Ser Ser Ser Gly Leu Ser Arg Met Gly Ala 365 370375 Val Pro Val Met Val Pro Ala Gln Ser Gln Ala Gly Ser Leu Val 380 385390 111 22 DNA Artificial Sequence Synthetic oligonucleotide probe 111agggtctcca ggagaaagac tc 22 112 24 DNA Artificial Sequence Syntheticoligonucleotide probe 112 attgtgggcc ttgcagacat agac 24 113 50 DNAArtificial Sequence Synthetic oligonucleotide probe 113 ggccacagcatcaaaacctt agaactcaat gtactggttc ctccagctcc 50 114 2479 DNA Homo Sapien114 acttgccatc acctgttgcc agtgtggaaa aattctccct gttgaatttt 50 ttgcacatggaggacagcag caaagagggc aacacaggct gataagacca 100 gagacagcag ggagattattttaccatacg ccctcaggac gttccctcta 150 gctggagttc tggacttcaa cagaaccccatccagtcatt ttgattttgc 200 tgtttatttt ttttttcttt ttctttttcc caccacattgtattttattt 250 ccgtacttca gaaatgggcc tacagaccac aaagtggccc agccatgggg300 cttttttcct gaagtcttgg cttatcattt ccctggggct ctactcacag 350gtgtccaaac tcctggcctg ccctagtgtg tgccgctgcg acaggaactt 400 tgtctactgtaatgagcgaa gcttgacctc agtgcctctt gggatcccgg 450 agggcgtaac cgtactctacctccacaaca accaaattaa taatgctgga 500 tttcctgcag aactgcacaa tgtacagtcggtgcacacgg tctacctgta 550 tggcaaccaa ctggacgaat tccccatgaa ccttcccaagaatgtcagag 600 ttctccattt gcaggaaaac aatattcaga ccatttcacg ggctgctctt650 gcccagctct tgaagcttga agagctgcac ctggatgaca actccatatc 700cacagtgggg gtggaagacg gggccttccg ggaggctatt agcctcaaat 750 tgttgtttttgtctaagaat cacctgagca gtgtgcctgt tgggcttcct 800 gtggacttgc aagagctgagagtggatgaa aatcgaattg ctgtcatatc 850 cgacatggcc ttccagaatc tcacgagcttggagcgtctt attgtggacg 900 ggaacctcct gaccaacaag ggtatcgccg agggcaccttcagccatctc 950 accaagctca aggaattttc aattgtacgt aattcgctgt cccaccctcc1000 tcccgatctc ccaggtacgc atctgatcag gctctatttg caggacaacc 1050agataaacca cattcctttg acagccttct caaatctgcg taagctggaa 1100 cggctggatatatccaacaa ccaactgcgg atgctgactc aaggggtttt 1150 tgataatctc tccaacctgaagcagctcac tgctcggaat aacccttggt 1200 tttgtgactg cagtattaaa tgggtcacagaatggctcaa atatatccct 1250 tcatctctca acgtgcgggg tttcatgtgc caaggtcctgaacaagtccg 1300 ggggatggcc gtcagggaat taaatatgaa tcttttgtcc tgtcccacca1350 cgacccccgg cctgcctctc ttcaccccag ccccaagtac agcttctccg 1400accactcagc ctcccaccct ctctattcca aaccctagca gaagctacac 1450 gcctccaactcctaccacat cgaaacttcc cacgattcct gactgggatg 1500 gcagagaaag agtgaccccacctatttctg aacggatcca gctctctatc 1550 cattttgtga atgatacttc cattcaagtcagctggctct ctctcttcac 1600 cgtgatggca tacaaactca catgggtgaa aatgggccacagtttagtag 1650 ggggcatcgt tcaggagcgc atagtcagcg gtgagaagca acacctgagc1700 ctggttaact tagagccccg atccacctat cggatttgtt tagtgccact 1750ggatgctttt aactaccgcg cggtagaaga caccatttgt tcagaggcca 1800 ccacccatgcctcctatctg aacaacggca gcaacacagc gtccagccat 1850 gagcagacga cgtcccacagcatgggctcc ccctttctgc tggcgggctt 1900 gatcgggggc gcggtgatat ttgtgctggtggtcttgctc agcgtctttt 1950 gctggcatat gcacaaaaag gggcgctaca cctcccagaagtggaaatac 2000 aaccggggcc ggcggaaaga tgattattgc gaggcaggca ccaagaagga2050 caactccatc ctggagatga cagaaaccag ttttcagatc gtctccttaa 2100ataacgatca actccttaaa ggagatttca gactgcagcc catttacacc 2150 ccaaatgggggcattaatta cacagactgc catatcccca acaacatgcg 2200 atactgcaac agcagcgtgccagacctgga gcactgccat acgtgacagc 2250 cagaggccca gcgttatcaa ggcggacaattagactcttg agaacacact 2300 cgtgtgtgca cataaagaca cgcagattac atttgataaatgttacacag 2350 atgcatttgt gcatttgaat actctgtaat ttatacggtg tactatataa2400 tgggatttaa aaaaagtgct atcttttcta tttcaagtta attacaaaca 2450gttttgtaac tctttgcttt ttaaatctt 2479 115 660 PRT Homo Sapien 115 Met GlyLeu Gln Thr Thr Lys Trp Pro Ser His Gly Ala Phe Phe 1 5 10 15 Leu LysSer Trp Leu Ile Ile Ser Leu Gly Leu Tyr Ser Gln Val 20 25 30 Ser Lys LeuLeu Ala Cys Pro Ser Val Cys Arg Cys Asp Arg Asn 35 40 45 Phe Val Tyr CysAsn Glu Arg Ser Leu Thr Ser Val Pro Leu Gly 50 55 60 Ile Pro Glu Gly ValThr Val Leu Tyr Leu His Asn Asn Gln Ile 65 70 75 Asn Asn Ala Gly Phe ProAla Glu Leu His Asn Val Gln Ser Val 80 85 90 His Thr Val Tyr Leu Tyr GlyAsn Gln Leu Asp Glu Phe Pro Met 95 100 105 Asn Leu Pro Lys Asn Val ArgVal Leu His Leu Gln Glu Asn Asn 110 115 120 Ile Gln Thr Ile Ser Arg AlaAla Leu Ala Gln Leu Leu Lys Leu 125 130 135 Glu Glu Leu His Leu Asp AspAsn Ser Ile Ser Thr Val Gly Val 140 145 150 Glu Asp Gly Ala Phe Arg GluAla Ile Ser Leu Lys Leu Leu Phe 155 160 165 Leu Ser Lys Asn His Leu SerSer Val Pro Val Gly Leu Pro Val 170 175 180 Asp Leu Gln Glu Leu Arg ValAsp Glu Asn Arg Ile Ala Val Ile 185 190 195 Ser Asp Met Ala Phe Gln AsnLeu Thr Ser Leu Glu Arg Leu Ile 200 205 210 Val Asp Gly Asn Leu Leu ThrAsn Lys Gly Ile Ala Glu Gly Thr 215 220 225 Phe Ser His Leu Thr Lys LeuLys Glu Phe Ser Ile Val Arg Asn 230 235 240 Ser Leu Ser His Pro Pro ProAsp Leu Pro Gly Thr His Leu Ile 245 250 255 Arg Leu Tyr Leu Gln Asp AsnGln Ile Asn His Ile Pro Leu Thr 260 265 270 Ala Phe Ser Asn Leu Arg LysLeu Glu Arg Leu Asp Ile Ser Asn 275 280 285 Asn Gln Leu Arg Met Leu ThrGln Gly Val Phe Asp Asn Leu Ser 290 295 300 Asn Leu Lys Gln Leu Thr AlaArg Asn Asn Pro Trp Phe Cys Asp 305 310 315 Cys Ser Ile Lys Trp Val ThrGlu Trp Leu Lys Tyr Ile Pro Ser 320 325 330 Ser Leu Asn Val Arg Gly PheMet Cys Gln Gly Pro Glu Gln Val 335 340 345 Arg Gly Met Ala Val Arg GluLeu Asn Met Asn Leu Leu Ser Cys 350 355 360 Pro Thr Thr Thr Pro Gly LeuPro Leu Phe Thr Pro Ala Pro Ser 365 370 375 Thr Ala Ser Pro Thr Thr GlnPro Pro Thr Leu Ser Ile Pro Asn 380 385 390 Pro Ser Arg Ser Tyr Thr ProPro Thr Pro Thr Thr Ser Lys Leu 395 400 405 Pro Thr Ile Pro Asp Trp AspGly Arg Glu Arg Val Thr Pro Pro 410 415 420 Ile Ser Glu Arg Ile Gln LeuSer Ile His Phe Val Asn Asp Thr 425 430 435 Ser Ile Gln Val Ser Trp LeuSer Leu Phe Thr Val Met Ala Tyr 440 445 450 Lys Leu Thr Trp Val Lys MetGly His Ser Leu Val Gly Gly Ile 455 460 465 Val Gln Glu Arg Ile Val SerGly Glu Lys Gln His Leu Ser Leu 470 475 480 Val Asn Leu Glu Pro Arg SerThr Tyr Arg Ile Cys Leu Val Pro 485 490 495 Leu Asp Ala Phe Asn Tyr ArgAla Val Glu Asp Thr Ile Cys Ser 500 505 510 Glu Ala Thr Thr His Ala SerTyr Leu Asn Asn Gly Ser Asn Thr 515 520 525 Ala Ser Ser His Glu Gln ThrThr Ser His Ser Met Gly Ser Pro 530 535 540 Phe Leu Leu Ala Gly Leu IleGly Gly Ala Val Ile Phe Val Leu 545 550 555 Val Val Leu Leu Ser Val PheCys Trp His Met His Lys Lys Gly 560 565 570 Arg Tyr Thr Ser Gln Lys TrpLys Tyr Asn Arg Gly Arg Arg Lys 575 580 585 Asp Asp Tyr Cys Glu Ala GlyThr Lys Lys Asp Asn Ser Ile Leu 590 595 600 Glu Met Thr Glu Thr Ser PheGln Ile Val Ser Leu Asn Asn Asp 605 610 615 Gln Leu Leu Lys Gly Asp PheArg Leu Gln Pro Ile Tyr Thr Pro 620 625 630 Asn Gly Gly Ile Asn Tyr ThrAsp Cys His Ile Pro Asn Asn Met 635 640 645 Arg Tyr Cys Asn Ser Ser ValPro Asp Leu Glu His Cys His Thr 650 655 660 116 21 DNA ArtificialSequence Synthetic oligonucleotide probe 116 cggtctacct gtatggcaac c 21117 22 DNA Artificial Sequence Synthetic oligonucleotide probe 117gcaggacaac cagataaacc ac 22 118 22 DNA Artificial Sequence Syntheticoligonucleotide probe 118 acgcagattt gagaaggctg tc 22 119 46 DNAArtificial Sequence Synthetic oligonucleotide probe 119 ttcacgggctgctcttgccc agctcttgaa gcttgaagag ctgcac 46 120 2857 DNA Homo Sapien 120tgaagagtaa tagttggaat caaaagagtc aacgcaatga actgttattt 50 actgctgcgttttatgttgg gaattcctct cctatggcct tgtcttggag 100 caacagaaaa ctctcaaacaaagaaagtca agcagccagt gcgatctcat 150 ttgagagtga agcgtggctg ggtgtggaaccaattttttg taccagagga 200 aatgaatacg actagtcatc acatcggcca gctaagatctgatttagaca 250 atggaaacaa ttctttccag tacaagcttt tgggagctgg agctggaagt300 acttttatca ttgatgaaag aacaggtgac atatatgcca tacagaagct 350tgatagagag gagcgatccc tctacatctt aagagcccag gtaatagaca 400 tcgctactggaagggctgtg gaacctgagt ctgagtttgt catcaaagtt 450 tcggatatca atgacaatgaaccaaaattc ctagatgaac cttatgaggc 500 cattgtacca gagatgtctc cagaaggaacattagttatc caggtgacag 550 caagtgatgc tgacgatccc tcaagtggta ataatgctcgtctcctctac 600 agcttacttc aaggccagcc atatttttct gttgaaccaa caacaggagt650 cataagaata tcttctaaaa tggatagaga actgcaagat gagtattggg 700taatcattca agccaaggac atgattggtc agccaggagc gttgtctgga 750 acaacaagtgtattaattaa actttcagat gttaatgaca ataagcctat 800 atttaaagaa agtttataccgcttgactgt ctctgaatct gcacccactg 850 ggacttctat aggaacaatc atggcatatgataatgacat aggagagaat 900 gcagaaatgg attacagcat tgaagaggat gattcgcaaacatttgacat 950 tattactaat catgaaactc aagaaggaat agttatatta aaaaagaaag1000 tggattttga gcaccagaac cactacggta ttagagcaaa agttaaaaac 1050catcatgttc ctgagcagct catgaagtac cacactgagg cttccaccac 1100 tttcattaagatccaggtgg aagatgttga tgagcctcct cttttcctcc 1150 ttccatatta tgtatttgaagtttttgaag aaaccccaca gggatcattt 1200 gtaggcgtgg tgtctgccac agacccagacaataggaaat ctcctatcag 1250 gtattctatt actaggagca aagtgttcaa tatcaatgataatggtacaa 1300 tcactacaag taactcactg gatcgtgaaa tcagtgcttg gtacaaccta1350 agtattacag ccacagaaaa atacaatata gaacagatct cttcgatccc 1400actgtatgtg caagttctta acatcaatga tcatgctcct gagttctctc 1450 aatactatgagacttatgtt tgtgaaaatg caggctctgg tcaggtaatt 1500 cagactatca gtgcagtggatagagatgaa tccatagaag agcaccattt 1550 ttactttaat ctatctgtag aagacactaacaattcaagt tttacaatca 1600 tagataatca agataacaca gctgtcattt tgactaatagaactggtttt 1650 aaccttcaag aagaacctgt cttctacatc tccatcttaa ttgccgacaa1700 tggaatcccg tcacttacaa gtacaaacac ccttaccatc catgtctgtg 1750actgtggtga cagtgggagc acacagacct gccagtacca ggagcttgtg 1800 ctttccatgggattcaagac agaagttatc attgctattc tcatttgcat 1850 tatgatcata tttgggtttatttttttgac tttgggttta aaacaacgga 1900 gaaaacagat tctatttcct gagaaaagtgaagatttcag agagaatata 1950 ttccaatatg atgatgaagg gggtggagaa gaagatacagaggcctttga 2000 tatagcagag ctgaggagta gtaccataat gcgggaacgc aagactcgga2050 aaaccacaag cgctgagatc aggagcctat acaggcagtc tttgcaagtt 2100ggccccgaca gtgccatatt caggaaattc attctggaaa agctcgaaga 2150 agctaatactgatccgtgtg cccctccttt tgattccctc cagacctacg 2200 cttttgaggg aacagggtcattagctggat ccctgagctc cttagaatca 2250 gcagtctctg atcaggatga aagctatgattaccttaatg agttgggacc 2300 tcgctttaaa agattagcat gcatgtttgg ttctgcagtgcagtcaaata 2350 attagggctt tttaccatca aaatttttaa aagtgctaat gtgtattcga2400 acccaatggt agtcttaaag agttttgtgc cctggctcta tggcggggaa 2450agccctagtc tatggagttt tctgatttcc ctggagtaaa tactccatgg 2500 ttattttaagctacctacat gctgtcattg aacagagatg tggggagaaa 2550 tgtaaacaat cagctcacaggcatcaatac aaccagattt gaagtaaaat 2600 aatgtaggaa gatattaaaa gtagatgagaggacacaaga tgtagtcgat 2650 ccttatgcga ttatatcatt atttacttag gaaagagtaaaaataccaaa 2700 cgagaaaatt taaaggagca aaaatttgca agtcaaatag aaatgtacaa2750 atcgagataa catttacatt tctatcatat tgacatgaaa attgaaaatg 2800tatagtcaga gaaattttca tgaattattc catgaagtat tgtttccttt 2850 atttaaa 2857121 772 PRT Homo Sapien 121 Met Asn Cys Tyr Leu Leu Leu Arg Phe Met LeuGly Ile Pro Leu 1 5 10 15 Leu Trp Pro Cys Leu Gly Ala Thr Glu Asn SerGln Thr Lys Lys 20 25 30 Val Lys Gln Pro Val Arg Ser His Leu Arg Val LysArg Gly Trp 35 40 45 Val Trp Asn Gln Phe Phe Val Pro Glu Glu Met Asn ThrThr Ser 50 55 60 His His Ile Gly Gln Leu Arg Ser Asp Leu Asp Asn Gly AsnAsn 65 70 75 Ser Phe Gln Tyr Lys Leu Leu Gly Ala Gly Ala Gly Ser Thr Phe80 85 90 Ile Ile Asp Glu Arg Thr Gly Asp Ile Tyr Ala Ile Gln Lys Leu 95100 105 Asp Arg Glu Glu Arg Ser Leu Tyr Ile Leu Arg Ala Gln Val Ile 110115 120 Asp Ile Ala Thr Gly Arg Ala Val Glu Pro Glu Ser Glu Phe Val 125130 135 Ile Lys Val Ser Asp Ile Asn Asp Asn Glu Pro Lys Phe Leu Asp 140145 150 Glu Pro Tyr Glu Ala Ile Val Pro Glu Met Ser Pro Glu Gly Thr 155160 165 Leu Val Ile Gln Val Thr Ala Ser Asp Ala Asp Asp Pro Ser Ser 170175 180 Gly Asn Asn Ala Arg Leu Leu Tyr Ser Leu Leu Gln Gly Gln Pro 185190 195 Tyr Phe Ser Val Glu Pro Thr Thr Gly Val Ile Arg Ile Ser Ser 200205 210 Lys Met Asp Arg Glu Leu Gln Asp Glu Tyr Trp Val Ile Ile Gln 215220 225 Ala Lys Asp Met Ile Gly Gln Pro Gly Ala Leu Ser Gly Thr Thr 230235 240 Ser Val Leu Ile Lys Leu Ser Asp Val Asn Asp Asn Lys Pro Ile 245250 255 Phe Lys Glu Ser Leu Tyr Arg Leu Thr Val Ser Glu Ser Ala Pro 260265 270 Thr Gly Thr Ser Ile Gly Thr Ile Met Ala Tyr Asp Asn Asp Ile 275280 285 Gly Glu Asn Ala Glu Met Asp Tyr Ser Ile Glu Glu Asp Asp Ser 290295 300 Gln Thr Phe Asp Ile Ile Thr Asn His Glu Thr Gln Glu Gly Ile 305310 315 Val Ile Leu Lys Lys Lys Val Asp Phe Glu His Gln Asn His Tyr 320325 330 Gly Ile Arg Ala Lys Val Lys Asn His His Val Pro Glu Gln Leu 335340 345 Met Lys Tyr His Thr Glu Ala Ser Thr Thr Phe Ile Lys Ile Gln 350355 360 Val Glu Asp Val Asp Glu Pro Pro Leu Phe Leu Leu Pro Tyr Tyr 365370 375 Val Phe Glu Val Phe Glu Glu Thr Pro Gln Gly Ser Phe Val Gly 380385 390 Val Val Ser Ala Thr Asp Pro Asp Asn Arg Lys Ser Pro Ile Arg 395400 405 Tyr Ser Ile Thr Arg Ser Lys Val Phe Asn Ile Asn Asp Asn Gly 410415 420 Thr Ile Thr Thr Ser Asn Ser Leu Asp Arg Glu Ile Ser Ala Trp 425430 435 Tyr Asn Leu Ser Ile Thr Ala Thr Glu Lys Tyr Asn Ile Glu Gln 440445 450 Ile Ser Ser Ile Pro Leu Tyr Val Gln Val Leu Asn Ile Asn Asp 455460 465 His Ala Pro Glu Phe Ser Gln Tyr Tyr Glu Thr Tyr Val Cys Glu 470475 480 Asn Ala Gly Ser Gly Gln Val Ile Gln Thr Ile Ser Ala Val Asp 485490 495 Arg Asp Glu Ser Ile Glu Glu His His Phe Tyr Phe Asn Leu Ser 500505 510 Val Glu Asp Thr Asn Asn Ser Ser Phe Thr Ile Ile Asp Asn Gln 515520 525 Asp Asn Thr Ala Val Ile Leu Thr Asn Arg Thr Gly Phe Asn Leu 530535 540 Gln Glu Glu Pro Val Phe Tyr Ile Ser Ile Leu Ile Ala Asp Asn 545550 555 Gly Ile Pro Ser Leu Thr Ser Thr Asn Thr Leu Thr Ile His Val 560565 570 Cys Asp Cys Gly Asp Ser Gly Ser Thr Gln Thr Cys Gln Tyr Gln 575580 585 Glu Leu Val Leu Ser Met Gly Phe Lys Thr Glu Val Ile Ile Ala 590595 600 Ile Leu Ile Cys Ile Met Ile Ile Phe Gly Phe Ile Phe Leu Thr 605610 615 Leu Gly Leu Lys Gln Arg Arg Lys Gln Ile Leu Phe Pro Glu Lys 620625 630 Ser Glu Asp Phe Arg Glu Asn Ile Phe Gln Tyr Asp Asp Glu Gly 635640 645 Gly Gly Glu Glu Asp Thr Glu Ala Phe Asp Ile Ala Glu Leu Arg 650655 660 Ser Ser Thr Ile Met Arg Glu Arg Lys Thr Arg Lys Thr Thr Ser 665670 675 Ala Glu Ile Arg Ser Leu Tyr Arg Gln Ser Leu Gln Val Gly Pro 680685 690 Asp Ser Ala Ile Phe Arg Lys Phe Ile Leu Glu Lys Leu Glu Glu 695700 705 Ala Asn Thr Asp Pro Cys Ala Pro Pro Phe Asp Ser Leu Gln Thr 710715 720 Tyr Ala Phe Glu Gly Thr Gly Ser Leu Ala Gly Ser Leu Ser Ser 725730 735 Leu Glu Ser Ala Val Ser Asp Gln Asp Glu Ser Tyr Asp Tyr Leu 740745 750 Asn Glu Leu Gly Pro Arg Phe Lys Arg Leu Ala Cys Met Phe Gly 755760 765 Ser Ala Val Gln Ser Asn Asn 770 122 25 DNA Artificial SequenceSynthetic oligonucleotide probe 122 cttgactgtc tctgaatctg caccc 25 12324 DNA Artificial Sequence Synthetic oligonucleotide probe 123aagtggtgga agcctccagt gtgg 24 124 52 DNA Artificial Sequence Syntheticoligonucleotide probe 124 ccactacggt attagagcaa aagttaaaaa ccatcatggttcctggagca 50 gc 52 125 1152 DNA Homo Sapien 125 cttcagaaca ggttctccttccccagtcac cagttgctcg agttagaatt 50 gtctgcaatg gccgccctgc agaaatctgtgagctctttc cttatgggga 100 ccctggccac cagctgcctc cttctcttgg ccctcttggtacagggagga 150 gcagctgcgc ccatcagctc ccactgcagg cttgacaagt ccaacttcca200 gcagccctat atcaccaacc gcaccttcat gctggctaag gaggctagct 250tggctgataa caacacagac gttcgtctca ttggggagaa actgttccac 300 ggagtcagtatgagtgagcg ctgctatctg atgaagcagg tgctgaactt 350 cacccttgaa gaagtgctgttccctcaatc tgataggttc cagccttata 400 tgcaggaggt ggtgcccttc ctggccaggctcagcaacag gctaagcaca 450 tgtcatattg aaggtgatga cctgcatatc cagaggaatgtgcaaaagct 500 gaaggacaca gtgaaaaagc ttggagagag tggagagatc aaagcaattg550 gagaactgga tttgctgttt atgtctctga gaaatgcctg catttgacca 600gagcaaagct gaaaaatgaa taactaaccc cctttccctg ctagaaataa 650 caattagatgccccaaagcg atttttttta accaaaagga agatgggaag 700 ccaaactcca tcatgatgggtggattccaa atgaacccct gcgttagtta 750 caaaggaaac caatgccact tttgtttataagaccagaag gtagactttc 800 taagcataga tatttattga taacatttca ttgtaactggtgttctatac 850 acagaaaaca atttattttt taaataattg tctttttcca taaaaaagat900 tactttccat tcctttaggg gaaaaaaccc ctaaatagct tcatgtttcc 950ataatcagta ctttatattt ataaatgtat ttattattat tataagactg 1000 cattttatttatatcatttt attaatatgg atttatttat agaaacatca 1050 ttcgatattg ctacttgagtgtaaggctaa tattgatatt tatgacaata 1100 attatagagc tataacatgt ttatttgacctcaataaaca cttggatatc 1150 cc 1152 126 179 PRT Homo Sapien 126 Met AlaAla Leu Gln Lys Ser Val Ser Ser Phe Leu Met Gly Thr 1 5 10 15 Leu AlaThr Ser Cys Leu Leu Leu Leu Ala Leu Leu Val Gln Gly 20 25 30 Gly Ala AlaAla Pro Ile Ser Ser His Cys Arg Leu Asp Lys Ser 35 40 45 Asn Phe Gln GlnPro Tyr Ile Thr Asn Arg Thr Phe Met Leu Ala 50 55 60 Lys Glu Ala Ser LeuAla Asp Asn Asn Thr Asp Val Arg Leu Ile 65 70 75 Gly Glu Lys Leu Phe HisGly Val Ser Met Ser Glu Arg Cys Tyr 80 85 90 Leu Met Lys Gln Val Leu AsnPhe Thr Leu Glu Glu Val Leu Phe 95 100 105 Pro Gln Ser Asp Arg Phe GlnPro Tyr Met Gln Glu Val Val Pro 110 115 120 Phe Leu Ala Arg Leu Ser AsnArg Leu Ser Thr Cys His Ile Glu 125 130 135 Gly Asp Asp Leu His Ile GlnArg Asn Val Gln Lys Leu Lys Asp 140 145 150 Thr Val Lys Lys Leu Gly GluSer Gly Glu Ile Lys Ala Ile Gly 155 160 165 Glu Leu Asp Leu Leu Phe MetSer Leu Arg Asn Ala Cys Ile 170 175 127 2557 DNA Homo Sapien 127gccctaacct tcccagggct cagctctttg gagctgccca ttcctccggc 50 tgcgagaaaggacgcgcgcc ctgcgtcggg cgaagaaaag aagcaaaact 100 tgtcgggagg gtttcgtcatcaacctcctt cccgcaaacc taaacctcct 150 gccggggcca tccctagaca gaggaaagttcctgcagagc cgaccagccc 200 tagtggatct ggggcaggca gcggcgctgg ctgtggaattagatctgttt 250 tgaacccagt ggagcgcatc gctggggctc ggaagtcacc gtccgcgggc300 accgggttgg cgctgcccga gtggaaccga cagtttgcga gcctcggctg 350caagtggcct ctcctccccg cggttgttgt tcagtgtcgg gtgagggctg 400 cgagtgtggcaagttgcaaa gagagcctca gaggtccgaa gagcgctgcg 450 ctcctactcg cgttcgcttcttcctcttct cggttcccta ctgtgaaatc 500 gcagcgacat ttacaaaggc ctccgggtcctaccgagacc gatccgcagc 550 gtttggcccg gtcgtgccta ttgcatcggg agcccccgagcaccggcgaa 600 atggcgaggt tcccgaaggc cgacctggcc gctgcaggag ttatgttact650 ttgccacttc ttcacggacc agtttcagtt cgccgatggg aaacccggag 700accaaatcct tgattggcag tatggagtta ctcaggcctt ccctcacaca 750 gaggaggaggtggaagttga ttcacacgcg tacagccaca ggtggaaaag 800 aaacttggac tttctcaaggcggtagacac gaaccgagca agcgtcggcc 850 aagactctcc tgagcccaga agcttcacagacctgctgct ggatgatggg 900 caggacaata acactcagat cgaggaggat acagaccacaattactatat 950 atctcgaata tatggtccat ctgattctgc cagccgggat ttatgggtga1000 acatagacca aatggaaaaa gataaagtga agattcatgg aatattgtcc 1050aatactcatc ggcaagctgc aagagtgaat ctgtccttcg attttccatt 1100 ttatggccacttcctacgtg aaatcactgt ggcaaccggg ggtttcatat 1150 acactggaga agtcgtacatcgaatgctaa cagccacaca gtacatagca 1200 cctttaatgg caaatttcga tcccagtgtatccagaaatt caactgtcag 1250 atattttgat aatggcacag cacttgtggt ccagtgggaccatgtacatc 1300 tccaggataa ttataacctg ggaagcttca cattccaggc aaccctgctc1350 atggatggac gaatcatctt tggatacaaa gaaattcctg tcttggtcac 1400acagataagt tcaaccaatc atccagtgaa agtcggactg tccgatgcat 1450 ttgtcgttgtccacaggatc caacaaattc ccaatgttcg aagaagaaca 1500 atttatgaat accaccgagtagagctacaa atgtcaaaaa ttaccaacat 1550 ttcggctgtg gagatgaccc cattacccacatgcctccag tttaacagat 1600 gtggcccctg tgtatcttct cagattggct tcaactgcagttggtgtagt 1650 aaacttcaaa gatgttccag tggatttgat cgtcatcggc aggactgggt1700 ggacagtgga tgccctgaag agtcaaaaga gaagatgtgt gagaatacag 1750aaccagtgga aacttcttct cgaaccacca caaccgtagg agcgacaacc 1800 acccagttcagggtcctaac taccaccaga agagcagtga cttctcagtt 1850 tcccaccagc ctccctacagaagatgatac caagatagca ctacatctaa 1900 aagataatgg agcttctaca gatgacagtgcagctgagaa gaaaggggga 1950 accctccacg ctggcctcat cattggaatc ctcatcctggtcctcattgt 2000 agccacagcc attcttgtga cagtctatat gtatcaccac ccaacatcag2050 cagccagcat cttctttatt gagagacgcc caagcagatg gcctgcgatg 2100aagtttagaa gaggctctgg acatcctgcc tatgctgaag ttgaaccagt 2150 tggagagaaagaaggcttta ttgtatcaga gcagtgctaa aatttctagg 2200 acagaacaac accagtactggtttacaggt gttaagacta aaattttgcc 2250 tataccttta agacaaacaa acaaacacacacacaaacaa gctctaagct 2300 gctgtagcct gaagaagaca agatttctgg acaagctcagcccaggaaac 2350 aaagggtaaa caaaaaacta aaacttatac aagataccat ttacactgaa2400 catagaattc cctagtggaa tgtcatctat agttcactcg gaacatctcc 2450cgtggactta tctgaagtat gacaagatta taatgctttt ggcttaggtg 2500 cagggttgcaaagggatcag aaaaaaaaaa tcataataaa gctttagttc 2550 atgaggg 2557 128 529PRT Homo Sapien 128 Met Ala Arg Phe Pro Lys Ala Asp Leu Ala Ala Ala GlyVal Met 1 5 10 15 Leu Leu Cys His Phe Phe Thr Asp Gln Phe Gln Phe AlaAsp Gly 20 25 30 Lys Pro Gly Asp Gln Ile Leu Asp Trp Gln Tyr Gly Val ThrGln 35 40 45 Ala Phe Pro His Thr Glu Glu Glu Val Glu Val Asp Ser His Ala50 55 60 Tyr Ser His Arg Trp Lys Arg Asn Leu Asp Phe Leu Lys Ala Val 6570 75 Asp Thr Asn Arg Ala Ser Val Gly Gln Asp Ser Pro Glu Pro Arg 80 8590 Ser Phe Thr Asp Leu Leu Leu Asp Asp Gly Gln Asp Asn Asn Thr 95 100105 Gln Ile Glu Glu Asp Thr Asp His Asn Tyr Tyr Ile Ser Arg Ile 110 115120 Tyr Gly Pro Ser Asp Ser Ala Ser Arg Asp Leu Trp Val Asn Ile 125 130135 Asp Gln Met Glu Lys Asp Lys Val Lys Ile His Gly Ile Leu Ser 140 145150 Asn Thr His Arg Gln Ala Ala Arg Val Asn Leu Ser Phe Asp Phe 155 160165 Pro Phe Tyr Gly His Phe Leu Arg Glu Ile Thr Val Ala Thr Gly 170 175180 Gly Phe Ile Tyr Thr Gly Glu Val Val His Arg Met Leu Thr Ala 185 190195 Thr Gln Tyr Ile Ala Pro Leu Met Ala Asn Phe Asp Pro Ser Val 200 205210 Ser Arg Asn Ser Thr Val Arg Tyr Phe Asp Asn Gly Thr Ala Leu 215 220225 Val Val Gln Trp Asp His Val His Leu Gln Asp Asn Tyr Asn Leu 230 235240 Gly Ser Phe Thr Phe Gln Ala Thr Leu Leu Met Asp Gly Arg Ile 245 250255 Ile Phe Gly Tyr Lys Glu Ile Pro Val Leu Val Thr Gln Ile Ser 260 265270 Ser Thr Asn His Pro Val Lys Val Gly Leu Ser Asp Ala Phe Val 275 280285 Val Val His Arg Ile Gln Gln Ile Pro Asn Val Arg Arg Arg Thr 290 295300 Ile Tyr Glu Tyr His Arg Val Glu Leu Gln Met Ser Lys Ile Thr 305 310315 Asn Ile Ser Ala Val Glu Met Thr Pro Leu Pro Thr Cys Leu Gln 320 325330 Phe Asn Arg Cys Gly Pro Cys Val Ser Ser Gln Ile Gly Phe Asn 335 340345 Cys Ser Trp Cys Ser Lys Leu Gln Arg Cys Ser Ser Gly Phe Asp 350 355360 Arg His Arg Gln Asp Trp Val Asp Ser Gly Cys Pro Glu Glu Ser 365 370375 Lys Glu Lys Met Cys Glu Asn Thr Glu Pro Val Glu Thr Ser Ser 380 385390 Arg Thr Thr Thr Thr Val Gly Ala Thr Thr Thr Gln Phe Arg Val 395 400405 Leu Thr Thr Thr Arg Arg Ala Val Thr Ser Gln Phe Pro Thr Ser 410 415420 Leu Pro Thr Glu Asp Asp Thr Lys Ile Ala Leu His Leu Lys Asp 425 430435 Asn Gly Ala Ser Thr Asp Asp Ser Ala Ala Glu Lys Lys Gly Gly 440 445450 Thr Leu His Ala Gly Leu Ile Ile Gly Ile Leu Ile Leu Val Leu 455 460465 Ile Val Ala Thr Ala Ile Leu Val Thr Val Tyr Met Tyr His His 470 475480 Pro Thr Ser Ala Ala Ser Ile Phe Phe Ile Glu Arg Arg Pro Ser 485 490495 Arg Trp Pro Ala Met Lys Phe Arg Arg Gly Ser Gly His Pro Ala 500 505510 Tyr Ala Glu Val Glu Pro Val Gly Glu Lys Glu Gly Phe Ile Val 515 520525 Ser Glu Gln Cys 129 4834 DNA Homo Sapien unsure 3784 unknown base129 gcagccctag cagggatgga catgatgctg ttggtgcagg gtgcttgttg 50 ctcgaaccagtggctggcgg cggtgctcct cagcctgtgc tgcctgctac 100 cctcctgcct cccggctggacagagtgtgg acttcccctg ggcggccgtg 150 gacaacatga tggtcagaaa aggggacacggcggtgctta ggtgttattt 200 ggaagatgga gcttcaaagg gtgcctggct gaaccggtcaagtattattt 250 ttgcgggagg tgataagtgg tcagtggatc ctcgagtttc aatttcaaca300 ttgaataaaa gggactacag cctccagata cagaatgtag atgtgacaga 350tgatggccca tacacgtgtt ctgttcagac tcaacataca cccagaacaa 400 tgcaggtgcatctaactgtg caagttcctc ctaagatata tgacatctca 450 aatgatatga ccgtcaatgaaggaaccaac gtcactctta cttgtttggc 500 cactgggaaa ccagagcctt ccatttcttggcgacacatc tccccatcag 550 caaaaccatt tgaaaatgga caatatttgg acatttatggaattacaagg 600 gaccaggctg gggaatatga atgcagtgcg gaaaatgatg tgtcattccc650 agatgtgagg aaagtaaaag ttgttgtcaa ctttgctcct actattcagg 700aaattaaatc tggcaccgtg acccccggac gcagtggcct gataagatgt 750 gaaggtgcaggtgtgccgcc tccagccttt gaatggtaca aaggagagaa 800 gaagctcttc aatggccaacaaggaattat tattcaaaat tttagcacaa 850 gatccattct cactgttacc aacgtgacacaggagcactt cggcaattat 900 acttgtgtgg ctgccaacaa gctaggcaca accaatgcgagcctgcctct 950 taaccctcca agtacagccc agtatggaat taccgggagc gctgatgttc1000 ttttctcctg ctggtacctt gtgttgacac tgtcctcttt caccagcata 1050ttctacctga agaatgccat tctacaataa attcaaagac ccataaaagg 1100 cttttaaggattctctgaaa gtgctgatgg ctggatccaa tctggtacag 1150 tttgttaaaa gcagcgtgggatataatcag cagtgcttac atggggatga 1200 tcgccttctg tagaattgct cattatgtaaatactttaat tctactcttt 1250 tttgattagc tacattacct tgtgaagcag tacacattgtccttttttta 1300 agacgtgaaa gctctgaaat tacttttaga ggatattaat tgtgatttca1350 tgtttgtaat ctacaacttt tcaaaagcat tcagtcatgg tctgctaggt 1400tgcaggctgt agtttacaaa aacgaatatt gcagtgaata tgtgattctt 1450 taaggctgcaatacaagcat tcagttccct gtttcaataa gagtcaatcc 1500 acatttacaa agatgcatttttttcttttt tgataaaaaa gcaaataata 1550 ttgccttcag attatttctt caaaatataacacatatcta gatttttctg 1600 ctcgcatgat attcaggttt caggaatgag ccttgtaatataactggctg 1650 tgcagctctg cttctctttc ctgtaagttc agcatgggtg tgccttcata1700 caataatatt tttctctttg tctccaacta atataaaatg ttttgctaaa 1750tcttacaatt tgaaagtaaa aataaaccag agtgatcaag ttaaaccata 1800 cactatctctaagtaacgaa ggagctattg gactgtaaaa atctcttcct 1850 gcactgacaa tggggtttgagaattttgcc ccacactaac tcagttcttg 1900 tgatgagaga caatttaata acagtatagtaaatatacca tatgatttct 1950 ttagttgtag ctaaatgtta gatccaccgt gggaaatcattccctttaaa 2000 atgacagcac agtccactca aaggattgcc tagcaataca gcatcttttc2050 ctttcactag tccaagccaa aaattttaag atgatttgtc agaaagggca 2100caaagtccta tcacctaata ttacaagagt tggtaagcgc tcatcattaa 2150 ttttattttgtggcagctaa gttagtatga cagaggcagt gctcctgtgg 2200 acaggagcat tttgcatattttccatctga aagtatcact cagttgatag 2250 tctggaatgc atgttatata ttttaaaacttccaaaatat attataacaa 2300 acattctata tcggtatgta gcagaccaat ctctaaaatagctaattctt 2350 caataaaatc tttctatata gccatttcag tgcaaacaag taaaatcaaa2400 aaagaccatc ctttattttt ccttacatga tatatgtaag atgcgatcaa 2450ataaagacaa aacaccagtg atgagaatat cttaagataa gtaattatca 2500 aattattgtgaatgttaaat tatttctact ataaagaagc aaaactacat 2550 ttttgaagga aaatgctgttactctaacat taatttacag gaatagtttg 2600 atggtttcac tctttactaa agaaaggccatcaccttgaa agccatttta 2650 caggtttgat gaagttacca atttcagtac acctaaatttctacaaatag 2700 tcccctttta caagttgtaa caacaaagac cctataataa aattagatac2750 aagaaatttt gcagtggtta tacatatttg agatatctag tatgttgccc 2800tagcagggat ggcttaaaaa ctgtgatttt ttttcttcaa gtaaaactta 2850 gtcccaaagtacatcataaa tcaattttaa ttagaaaaat gaatcttaaa 2900 tgaggggaca taagtatactctttccacaa aatggcaata ataaggcata 2950 aagctagtaa atctactaac tgtaataaatgtatgacatt attttgattg 3000 atacattaaa aaagagtttt tagaacaaat atggcatttaactttattat 3050 ttatttgctt ttaagaaata ttctttgtgg aattgttgaa taaactataa3100 aatattattt tgtattgcag ctttaaagtg gcacactcca taataatcta 3150cttactagaa atagtggtgc taccacaaaa aatgttaacc atcagtacca 3200 ttgtttgggagaaagaaaca gatcaagaat gcatattatt cagtgaccgc 3250 tttcctagag ttaaaatacctcctctttgt aaggtttgta ggtaaattga 3300 ggtataaact atggatgaac caaataattagttcaaagtg ttgtcatgat 3350 tccaaatttg tggagtctgg tgtttttacc atagaatgtgacagaagtac 3400 agtcatagct cagtagctat atgtatttgc ctttatgtta gaagagactt3450 tcttgagtga catttttaaa tagaggaggt attcactatg tttttctgta 3500tcacagcagc attcctagtc cttaggccct cggacagagt gaaatcatga 3550 gtatttatgagttcaatatt gtcaaataag gctacagtat ttgctttttt 3600 gtgtgaatgt attgcatataatgttcaagt agatgatttt acatttatgg 3650 acatataaaa tgtctgatta ccccattttatcagtcctga ctgtacaaga 3700 ttgttgcaat ttcagaatag cagttttata aattgatttatcttttaatc 3750 tataacaatt tgtgttagct gttcatttca ggantatatt ttctacaagt3800 tccacttgtg ggactccttt tgttgcccct attttttttt aaagaaggaa 3850gaaagaaaaa taagtagcag tttaaaaatg agaatggaga gaaaagaaaa 3900 agaatgaaaaggaaaggcag taaagaggga aaaaaaagga aggatggaag 3950 gaatgaagga aggaagggaggaaggggaga aggtaggaag aaagaaagga 4000 tgagagggaa ggaagaatca gagtattagggtagttaact tacacatttg 4050 cattcttagt ttaactgcaa gtggtgtaac tatgtttttcaatgatcgca 4100 tttgaaacat aagtcctatt ataccattaa gttcctatta tgcagcaatt4150 atataataaa aagtactgcc caagttatag taatgtgggt gtttttgaga 4200cactaaaaga tttgagaggg agaatttcaa acttaaagcc acttttgggg 4250 ggtttataacttaactgaaa aattaatgct tcatcataac atttaagcta 4300 tatctagaaa gtagactggagaactgagaa aattacccag gtaattcagg 4350 gaaaaaaaaa aatatatata tatataaatacccctacatt tgaagtcaga 4400 aaactctgaa aaactgaatt atcaaagtca atcatctataatgatcaaat 4450 ttactgaaca attgttaatt tatccattgt gcttagcttt gtgacacagc4500 caaaagttac ctatttaatc ttttcaataa aaattgtttt ttgaaatcca 4550gaaatgattt aaaaagaggt caggttttta actatttatt gaagtatgtg 4600 gatgtacagtatttcaatag atatgaatat gaataaatgg tatgccttaa 4650 gattctttga atatgtatttactttaaaga ctggaaaaag ctcttcctgt 4700 cttttagtaa aacatccata tttcataacctgatgtaaaa tatgttgtac 4750 tgtttccaat aggtgaatat aaactcagtt tatcaattaaaaaaaaaaaa 4800 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaa 4834 130 354 PRTHomo Sapien 130 Met Asp Met Met Leu Leu Val Gln Gly Ala Cys Cys Ser AsnGln 1 5 10 15 Trp Leu Ala Ala Val Leu Leu Ser Leu Cys Cys Leu Leu ProSer 20 25 30 Cys Leu Pro Ala Gly Gln Ser Val Asp Phe Pro Trp Ala Ala Val35 40 45 Asp Asn Met Met Val Arg Lys Gly Asp Thr Ala Val Leu Arg Cys 5055 60 Tyr Leu Glu Asp Gly Ala Ser Lys Gly Ala Trp Leu Asn Arg Ser 65 7075 Ser Ile Ile Phe Ala Gly Gly Asp Lys Trp Ser Val Asp Pro Arg 80 85 90Val Ser Ile Ser Thr Leu Asn Lys Arg Asp Tyr Ser Leu Gln Ile 95 100 105Gln Asn Val Asp Val Thr Asp Asp Gly Pro Tyr Thr Cys Ser Val 110 115 120Gln Thr Gln His Thr Pro Arg Thr Met Gln Val His Leu Thr Val 125 130 135Gln Val Pro Pro Lys Ile Tyr Asp Ile Ser Asn Asp Met Thr Val 140 145 150Asn Glu Gly Thr Asn Val Thr Leu Thr Cys Leu Ala Thr Gly Lys 155 160 165Pro Glu Pro Ser Ile Ser Trp Arg His Ile Ser Pro Ser Ala Lys 170 175 180Pro Phe Glu Asn Gly Gln Tyr Leu Asp Ile Tyr Gly Ile Thr Arg 185 190 195Asp Gln Ala Gly Glu Tyr Glu Cys Ser Ala Glu Asn Asp Val Ser 200 205 210Phe Pro Asp Val Arg Lys Val Lys Val Val Val Asn Phe Ala Pro 215 220 225Thr Ile Gln Glu Ile Lys Ser Gly Thr Val Thr Pro Gly Arg Ser 230 235 240Gly Leu Ile Arg Cys Glu Gly Ala Gly Val Pro Pro Pro Ala Phe 245 250 255Glu Trp Tyr Lys Gly Glu Lys Lys Leu Phe Asn Gly Gln Gln Gly 260 265 270Ile Ile Ile Gln Asn Phe Ser Thr Arg Ser Ile Leu Thr Val Thr 275 280 285Asn Val Thr Gln Glu His Phe Gly Asn Tyr Thr Cys Val Ala Ala 290 295 300Asn Lys Leu Gly Thr Thr Asn Ala Ser Leu Pro Leu Asn Pro Pro 305 310 315Ser Thr Ala Gln Tyr Gly Ile Thr Gly Ser Ala Asp Val Leu Phe 320 325 330Ser Cys Trp Tyr Leu Val Leu Thr Leu Ser Ser Phe Thr Ser Ile 335 340 345Phe Tyr Leu Lys Asn Ala Ile Leu Gln 350 131 823 DNA Homo Sapien 131atagtagaag aatgtctctg aaattactgg atgagtttca gtcatacttt 50 cacatgggcacaatttcaca ttcaagctcc ttatcctagg ctaattttat 100 attatgttaa atcacttgtttttgttctca cggcttcctg cctgctatag 150 gcataattac gaggaagcag aacttctccagaagcaagcg cacatgcgtt 200 ccaaaataag agcaaattcg ctctaaacac aggaaaagacctgaagcttt 250 aattaagggg ttacatccaa ccccagagcg cttttgtggg cactgattgc300 tccagcttct gcgtcactgc gcgagggaag agggaagagg atccaggcgt 350tagacatgta tagacacaaa aacagctgga gattgggctt aaaataccca 400 ccaagctccaaagaagagac ccaagtcccc aaaacattga tttcagggct 450 gccaggaagg aagagcagcagcagggtggg agagaagctc cagtcagccc 500 acaagatgcc attgtccccc ggcctcctgctgctgctgct ctccggggcc 550 acggccaccg ctgccctgcc cctggagggt ggccccaccggccgagacag 600 cgagcatatg caggaagcgg caggaataag gaaaagcagc ctcctgactt650 tcctcgcttg gtggtttgag tggacctccc aggccagtgc cgggcccctc 700ataggagagg aagctcggga ggtggccagg cggcaggaag gcgcaccccc 750 ccagcaatccgcgcgccggg acagaatgcc ctgcaggaac ttcttctgga 800 agaccttctc ctcctgcaaatag 823 132 155 PRT Homo Sapien 132 Met Tyr Arg His Lys Asn Ser Trp ArgLeu Gly Leu Lys Tyr Pro 1 5 10 15 Pro Ser Ser Lys Glu Glu Thr Gln ValPro Lys Thr Leu Ile Ser 20 25 30 Gly Leu Pro Gly Arg Lys Ser Ser Ser ArgVal Gly Glu Lys Leu 35 40 45 Gln Ser Ala His Lys Met Pro Leu Ser Pro GlyLeu Leu Leu Leu 50 55 60 Leu Leu Ser Gly Ala Thr Ala Thr Ala Ala Leu ProLeu Glu Gly 65 70 75 Gly Pro Thr Gly Arg Asp Ser Glu His Met Gln Glu AlaAla Gly 80 85 90 Ile Arg Lys Ser Ser Leu Leu Thr Phe Leu Ala Trp Trp PheGlu 95 100 105 Trp Thr Ser Gln Ala Ser Ala Gly Pro Leu Ile Gly Glu GluAla 110 115 120 Arg Glu Val Ala Arg Arg Gln Glu Gly Ala Pro Pro Gln GlnSer 125 130 135 Ala Arg Arg Asp Arg Met Pro Cys Arg Asn Phe Phe Trp LysThr 140 145 150 Phe Ser Ser Cys Lys 155 133 24 DNA Artificial SequenceSynthetic oligonucleotide probe 133 tcagggctgc caggaaggaa gagc 24 134 28DNA Artificial Sequence Synthetic oligonucleotide probe 134 gcaggaggagaaggtcttcc agaagaag 28 135 45 DNA Artificial Sequence Syntheticoligonucleotide probe 135 agaagttcca gtcagcccac aagatgccat tgtcccccggcctcc 45 136 1875 DNA Homo Sapien 136 gtcgtgtgct tggaggaagc cgcggaacccccagcgtccg tccatggcgt 50 ggagccttgg gagctggctg ggtggctgcc tgctggtgtcagcattggga 100 atggtaccac ctcccgaaaa tgtcagaatg aattctgtta atttcaagaa150 cattctacag tgggagtcac ctgcttttgc caaagggaac ctgactttca 200cagctcagta cctaagttat aggatattcc aagataaatg catgaatact 250 accttgacggaatgtgattt ctcaagtctt tccaagtatg gtgaccacac 300 cttgagagtc agggctgaatttgcagatga gcattcagac tgggtaaaca 350 tcaccttctg tcctgtggat gacaccattattggaccccc tggaatgcaa 400 gtagaagtac ttgctgattc tttacatatg cgtttcttagcccctaaaat 450 tgagaatgaa tacgaaactt ggactatgaa gaatgtgtat aactcatgga500 cttataatgt gcaatactgg aaaaacggta ctgatgaaaa gtttcaaatt 550actccccagt atgactttga ggtcctcaga aacctggagc catggacaac 600 ttattgtgttcaagttcgag ggtttcttcc tgatcggaac aaagctgggg 650 aatggagtga gcctgtctgtgagcaaacaa cccatgacga aacggtcccc 700 tcctggatgg tggccgtcat cctcatggcctcggtcttca tggtctgcct 750 ggcactcctc ggctgcttct ccttgctgtg gtgcgtttacaagaagacaa 800 agtacgcctt ctcccctagg aattctcttc cacagcacct gaaagagttt850 ttgggccatc ctcatcataa cacacttctg tttttctcct ttccattgtc 900ggatgagaat gatgtttttg acaagctaag tgtcattgca gaagactctg 950 agagcggcaagcagaatcct ggtgacagct gcagcctcgg gaccccgcct 1000 gggcaggggc cccaaagctaggctctgaga aggaaacaca ctcggctggg 1050 cacagtgacg tactccatct cacatctgcctcagtgaggg atcagggcag 1100 caaacaaggg ccaagaccat ctgagccagc cccacatctagaactccaga 1150 cctggactta gccaccagag agctacattt taaaggctgt cttggcaaaa1200 atactccatt tgggaactca ctgccttata aaggctttca tgatgttttc 1250agaagttggc cactgagagt gtaattttca gccttttata tcactaaaat 1300 aagatcatgttttaattgtg agaaacaggg ccgagcacag tggctcacgc 1350 ctgtaatacc agcaccttagaggtcgaggc aggcggatca cttgaggtca 1400 ggagttcaag accagcctgg ccaatatggtgaaacccagt ctctactaaa 1450 aatacaaaaa ttagctaggc atgatggcgc atgcctataatcccagctac 1500 tcgagtgcct gaggcaggag aattgcatga acccgggagg aggaggagga1550 ggttgcagtg agccgagata gcggcactgc actccagcct gggtgacaaa 1600gtgagactcc atctcaaaaa aaaaaaaaaa aaattgtgag aaacagaaat 1650 acttaaaatgaggaataaga atggagatgt tacatctggt agatgtaaca 1700 ttctaccaga ttatggatggactgatctga aaatcgacct caactcaagg 1750 gtggtcagct caatgctaca cagagcacggacttttggat tctttgcagt 1800 actttgaatt tatttttcta cctatatatg ttttatatgctgctggtgct 1850 ccattaaagt tttactctgt gttgc 1875 137 325 PRT Homo Sapien137 Met Ala Trp Ser Leu Gly Ser Trp Leu Gly Gly Cys Leu Leu Val 1 5 1015 Ser Ala Leu Gly Met Val Pro Pro Pro Glu Asn Val Arg Met Asn 20 25 30Ser Val Asn Phe Lys Asn Ile Leu Gln Trp Glu Ser Pro Ala Phe 35 40 45 AlaLys Gly Asn Leu Thr Phe Thr Ala Gln Tyr Leu Ser Tyr Arg 50 55 60 Ile PheGln Asp Lys Cys Met Asn Thr Thr Leu Thr Glu Cys Asp 65 70 75 Phe Ser SerLeu Ser Lys Tyr Gly Asp His Thr Leu Arg Val Arg 80 85 90 Ala Glu Phe AlaAsp Glu His Ser Asp Trp Val Asn Ile Thr Phe 95 100 105 Cys Pro Val AspAsp Thr Ile Ile Gly Pro Pro Gly Met Gln Val 110 115 120 Glu Val Leu AlaAsp Ser Leu His Met Arg Phe Leu Ala Pro Lys 125 130 135 Ile Glu Asn GluTyr Glu Thr Trp Thr Met Lys Asn Val Tyr Asn 140 145 150 Ser Trp Thr TyrAsn Val Gln Tyr Trp Lys Asn Gly Thr Asp Glu 155 160 165 Lys Phe Gln IleThr Pro Gln Tyr Asp Phe Glu Val Leu Arg Asn 170 175 180 Leu Glu Pro TrpThr Thr Tyr Cys Val Gln Val Arg Gly Phe Leu 185 190 195 Pro Asp Arg AsnLys Ala Gly Glu Trp Ser Glu Pro Val Cys Glu 200 205 210 Gln Thr Thr HisAsp Glu Thr Val Pro Ser Trp Met Val Ala Val 215 220 225 Ile Leu Met AlaSer Val Phe Met Val Cys Leu Ala Leu Leu Gly 230 235 240 Cys Phe Ser LeuLeu Trp Cys Val Tyr Lys Lys Thr Lys Tyr Ala 245 250 255 Phe Ser Pro ArgAsn Ser Leu Pro Gln His Leu Lys Glu Phe Leu 260 265 270 Gly His Pro HisHis Asn Thr Leu Leu Phe Phe Ser Phe Pro Leu 275 280 285 Ser Asp Glu AsnAsp Val Phe Asp Lys Leu Ser Val Ile Ala Glu 290 295 300 Asp Ser Glu SerGly Lys Gln Asn Pro Gly Asp Ser Cys Ser Leu 305 310 315 Gly Thr Pro ProGly Gln Gly Pro Gln Ser 320 325 138 2570 DNA Homo Sapien 138 cgagcgccaacccgctagcg cctgaatccg gcgtgctgcc cgctcgccgc 50 ccgccatggc ccgcgcagccccgctgctcg ccgcgttgac cgcgctcctc 100 gccgccgccg ctgctggcgg agatgccccgccgggcaaaa tcgcggtggt 150 tggggctggg attgggggct ctgctgtggc ccattttctccagcagcact 200 ttggacctcg ggtgcagatc gacgtgtacg agaagggaac cgtgggtggc250 cgcttggcca ccatctcagt caacaagcag cactatgaga gcggggctgc 300ctccttccac tccctgagcc tgcacatgca ggacttcgtc aagctgctgg 350 ggctgaggcaccggcgcgag gtggtgggca ggagcgccat cttcggcggg 400 gagcacttca tgctggaggagactgactgg tacctgctga acctcttccg 450 cctctggtgg cactatggca tcagcttcctgaggctgcag atgtgggtgg 500 aggaggtcat ggagaagttc atgaggatct ataagtaccaggcccacggc 550 tatgccttct cgggtgtgga ggagctgctc tactcactgg gggagtccac600 ctttgttaac atgacccagc actctgtggc tgagtccctg ctgcaggtgg 650gcgtcacgca gcgctttatt gatgatgtcg tttctgctgt cctgcgggcc 700 agctatggccagtcagcagc gatgcccgcc tttgcaggag ccatgtcact 750 agccggggcc caaggcagcctgtggtctgt ggaaggaggc aataagctgg 800 tttgttccgg tttgctgaag ctcaccaaggccaatgtgat ccatgccaca 850 gtgacctctg tgaccctgca cagcacagag gggaaagccctgtaccaggt 900 ggcgtatgag aatgaggtag gcaacagctc tgacttctat gacatcgtgg950 tcatcgccac ccccctgcac ctggacaaca gcagcagcaa cttaaccttt 1000gcaggcttcc acccgcccat tgatgacgtg cagggctctt tccagcccac 1050 cgtcgtctccttggtccacg gctacctcaa ctcgtcctac ttcggtttcc 1100 cagaccctaa gcttttcccctttgccaaca tccttaccac agatttcccc 1150 agcttcttct gcactctgga caacatctgccctgtcaaca tctctgccag 1200 cttccggcga aagcagcccc aggaggcagc tgtttggcgagtccagtccc 1250 ccaagcccct ctttcggacc cagctaaaga ccctgttccg ttcctattac1300 tcagtgcaga cagctgagtg gcaggcccat cccctctatg gctcccgccc 1350cacgctcccg aggtttgcac tccatgacca gctcttctac ctcaatgccc 1400 tggagtgggcggccagctcc gtggaggtga tggccgtggc tgccaagaat 1450 gtggccttgc tggcttacaaccgctggtac caggacctag acaagattga 1500 tcaaaaagat ttgatgcaca aggtcaagactgaactgtga gggctctagg 1550 gagagcctgg gaactttcat cccccactga agatggatcatcccacagca 1600 gcccaggact gaataagcca tgctcgccca ccaggcttct ttctgacccc1650 tcatgtatca agcatctcca ggtgacctac tgtctgccta tattaagggt 1700ccacacggcg gctgctgctt ttttttaagg gggaaagtaa gaaaagagaa 1750 ggaaatccaagccagtatat ttgttttatt tatttttttt aagaagaaaa 1800 aagttcatct tcacaaggtgcttcagactt ggtttcttag ctagaaacca 1850 gaagactacg ggagggaata taaggcagagaactatgagt cttattttat 1900 tactgttttt cactacctac tcccacaatg gacaatcaattgaggcaacc 1950 tacaagaaaa catttacaac cagatggtta caaataaagt agaagggaag2000 atcagaaaac ctaagaaatg atcatagctc ctggttactg tggacttgat 2050ggatttgaag tacctagttc agaactccct agtcaccatc tccaagcctg 2100 tcaacatcactgcatattgg aggagatgac tgtggtagga cccaaggaag 2150 agatgtgtgc ctgaatagtcgtcaccatat ctccaagctt cctggcaacc 2200 agtgggaaaa gaaacatgcg aggctgtaggaagagggaag ctcttccttg 2250 gcacctagag gaattagcca ttctcttcct tatgcaaagattgaggaatg 2300 caacaatata aagaagagaa gtccccagat ggtagagagc agtcatatct2350 tacccctaga tgttcatccc agcagaagaa agaagaaggt gttggggtag 2400gattcttcag aggttagcct ggtactttct catcagacac tagcttgaag 2450 taagaggagaattatgcttt tctttgcttt ttctacaaac ccttaaaaat 2500 cacttgtttt aaaaagaaagtaaaagccct tttcattcaa aaaaaaaaaa 2550 aaaaaaaaaa aaaaaaaaaa 2570 139 494PRT Homo Sapien 139 Met Ala Arg Ala Ala Pro Leu Leu Ala Ala Leu Thr AlaLeu Leu 1 5 10 15 Ala Ala Ala Ala Ala Gly Gly Asp Ala Pro Pro Gly LysIle Ala 20 25 30 Val Val Gly Ala Gly Ile Gly Gly Ser Ala Val Ala His PheLeu 35 40 45 Gln Gln His Phe Gly Pro Arg Val Gln Ile Asp Val Tyr Glu Lys50 55 60 Gly Thr Val Gly Gly Arg Leu Ala Thr Ile Ser Val Asn Lys Gln 6570 75 His Tyr Glu Ser Gly Ala Ala Ser Phe His Ser Leu Ser Leu His 80 8590 Met Gln Asp Phe Val Lys Leu Leu Gly Leu Arg His Arg Arg Glu 95 100105 Val Val Gly Arg Ser Ala Ile Phe Gly Gly Glu His Phe Met Leu 110 115120 Glu Glu Thr Asp Trp Tyr Leu Leu Asn Leu Phe Arg Leu Trp Trp 125 130135 His Tyr Gly Ile Ser Phe Leu Arg Leu Gln Met Trp Val Glu Glu 140 145150 Val Met Glu Lys Phe Met Arg Ile Tyr Lys Tyr Gln Ala His Gly 155 160165 Tyr Ala Phe Ser Gly Val Glu Glu Leu Leu Tyr Ser Leu Gly Glu 170 175180 Ser Thr Phe Val Asn Met Thr Gln His Ser Val Ala Glu Ser Leu 185 190195 Leu Gln Val Gly Val Thr Gln Arg Phe Ile Asp Asp Val Val Ser 200 205210 Ala Val Leu Arg Ala Ser Tyr Gly Gln Ser Ala Ala Met Pro Ala 215 220225 Phe Ala Gly Ala Met Ser Leu Ala Gly Ala Gln Gly Ser Leu Trp 230 235240 Ser Val Glu Gly Gly Asn Lys Leu Val Cys Ser Gly Leu Leu Lys 245 250255 Leu Thr Lys Ala Asn Val Ile His Ala Thr Val Thr Ser Val Thr 260 265270 Leu His Ser Thr Glu Gly Lys Ala Leu Tyr Gln Val Ala Tyr Glu 275 280285 Asn Glu Val Gly Asn Ser Ser Asp Phe Tyr Asp Ile Val Val Ile 290 295300 Ala Thr Pro Leu His Leu Asp Asn Ser Ser Ser Asn Leu Thr Phe 305 310315 Ala Gly Phe His Pro Pro Ile Asp Asp Val Gln Gly Ser Phe Gln 320 325330 Pro Thr Val Val Ser Leu Val His Gly Tyr Leu Asn Ser Ser Tyr 335 340345 Phe Gly Phe Pro Asp Pro Lys Leu Phe Pro Phe Ala Asn Ile Leu 350 355360 Thr Thr Asp Phe Pro Ser Phe Phe Cys Thr Leu Asp Asn Ile Cys 365 370375 Pro Val Asn Ile Ser Ala Ser Phe Arg Arg Lys Gln Pro Gln Glu 380 385390 Ala Ala Val Trp Arg Val Gln Ser Pro Lys Pro Leu Phe Arg Thr 395 400405 Gln Leu Lys Thr Leu Phe Arg Ser Tyr Tyr Ser Val Gln Thr Ala 410 415420 Glu Trp Gln Ala His Pro Leu Tyr Gly Ser Arg Pro Thr Leu Pro 425 430435 Arg Phe Ala Leu His Asp Gln Leu Phe Tyr Leu Asn Ala Leu Glu 440 445450 Trp Ala Ala Ser Ser Val Glu Val Met Ala Val Ala Ala Lys Asn 455 460465 Val Ala Leu Leu Ala Tyr Asn Arg Trp Tyr Gln Asp Leu Asp Lys 470 475480 Ile Asp Gln Lys Asp Leu Met His Lys Val Lys Thr Glu Leu 485 490 14023 DNA Artificial Sequence Synthetic oligonucleotide probe 140gggacgtgct tctacaagaa cag 23 141 26 DNA Artificial Sequence Syntheticoligonucleotide probe 141 caggcttaca atgttatgat cagaca 26 142 31 DNAArtificial Sequence Synthetic oligonucleotide probe 142 tattcagagttttccattgg cagtgccagt t 31 143 18 DNA Artificial Sequence Syntheticoligonucleotide probe 143 ggccttgcag acaaccgt 18 144 21 DNA ArtificialSequence Synthetic oligonucleotide probe 144 cagactgagg gagatccgag a 21145 26 DNA Artificial Sequence Synthetic oligonucleotide probe 145gcagattttg aggacagcca cctcca 26 146 18 DNA Artificial Sequence Syntheticoligonucleotide probe 146 catcaagcgc ctctacca 18 147 21 DNA ArtificialSequence Synthetic oligonucleotide probe 147 cacaaactcg aactgcttct g 21148 20 DNA Artificial Sequence Synthetic oligonucleotide probe 148cagctgccct tccccaacca 20 149 22 DNA Artificial Sequence Syntheticoligonucleotide probe 149 ggcagagact tccagtcact ga 22 150 22 DNAArtificial Sequence Synthetic oligonucleotide probe 150 gccaagggtggtgttagata gg 22 151 24 DNA Artificial Sequence Syntheticoligonucleotide probe 151 caggccccct tgatctgtac ccca 24

What is claimed is:
 1. Isolated nucleic acid having at least 80% nucleicacid sequence identity to a nucleotide sequence that encodes an aminoacid sequence selected from the group consisting of the amino acidsequence shown in FIG. 2 (SEQ ID NO: 4), FIG. 4 (SEQ ID NO: 9), FIG. 6(SEQ ID NO: 11), FIG. 8 (SEQ ID NO: 13), FIG. 10 (SEQ ID NO: 15), FIG.12 (SEQ ID NO: 17), FIG. 14 (SEQ ID NO: 22), FIG. 16 (SEQ ID NO: 24),FIG. 18 (SEQ ID NO: 29), FIG. 20 (SEQ ID NO: 32), FIG. 22 (SEQ ID NO:39), FIG. 24 (SEQ ID NO: 41), FIG. 26 (SEQ ID NO: 52), FIG. 28 (SEQ IDNO: 54), FIG. 30 (SEQ ID NO: 56), FIG. 32 (SEQ ID NO: 58), FIG. 34 (SEQID NO: 63), FIG. 36 (SEQ ID NO: 65), FIG. 38 (SEQ ID NO: 73), FIG. 40(SEQ ID NO: 78), FIG. 42 (SEQ ID NO: 80), FIG. 44 (SEQ ID NO: 84), FIG.46 (SEQ ID NO: 86), FIG. 48 (SEQ ID NO: 91), FIG. 50 (SEQ ID NO: 99),FIG. 52 (SEQ ID NO: 104), FIG. 54 (SEQ ID NO: 106), FIG. 56 (SEQ ID NO:108), FIG. 58 (SEQ ID NO: 110), FIG. 60 (SEQ ID NO: 115), FIG. 62 (SEQID NO: 121), FIG. 64 (SEQ ID NO: 126), FIG. 66 (SEQ ID NO: 128), FIG. 68(SEQ ID NO: 130), FIG. 70 (SEQ ID NO: 132), FIG. 72 (SEQ ID NO: 137) andFIG. 74 (SEQ ID NO: 139).
 2. Isolated nucleic acid having at least 80%nucleic acid sequence identity to a nucleotide sequence selected fromthe group consisting of the nucleotide sequence shown in FIG. 1 (SEQ IDNO: 3), FIG. 3 (SEQ ID NO: 8), FIG. 5 (SEQ ID NO: 10), FIG. 7 (SEQ IDNO: 12), FIG. 9 (SEQ ID NO: 14), FIG. 11 (SEQ ID NO: 16), FIG. 13 (SEQID NO: 21), FIG. 15 (SEQ ID NO: 23), FIG. 17 (SEQ ID NO: 28), FIG. 19(SEQ ID NO: 31), FIG. 21 (SEQ ID NO: 38), FIG. 23 (SEQ ID NO: 40), FIG.25 (SEQ ID NO: 51), FIG. 27 (SEQ ID NO: 53), FIG. 29 (SEQ ID NO: 55),FIG. 31 (SEQ ID NO: 57), FIG. 33 (SEQ ID NO: 62), FIG. 35 (SEQ ID NO:64), FIG. 37 (SEQ ID NO: 72), FIG. 39 (SEQ ID NO: 77), FIG. 41 (SEQ IDNO: 79), FIG. 43 (SEQ ID NO: 83), FIG. 45 (SEQ ID NO: 85), FIG. 47 (SEQID NO: 90), FIG. 49 (SEQ ID NO: 98), FIG. 51 (SEQ ID NO: 103), FIG. 53(SEQ ID NO: 105), FIG. 55 (SEQ ID NO: 107), FIG. 57 (SEQ ID NO: 109),FIG. 59 (SEQ ID NO: 114), FIG. 61 (SEQ ID NO: 120), FIG. 63 (SEQ ID NO:125), FIG. 65 (SEQ ID NO: 127), FIGS. 67A-B (SEQ ID NO: 129), FIG. 69(SEQ ID NO: 131), FIG. 71 (SEQ ID NO: 136) and FIG. 73 (SEQ ID NO: 138).3. Isolated nucleic acid having at least 80% nucleic acid sequenceidentity to a nucleotide sequence selected from the group consisting ofthe full-length coding sequence of the nucleotide sequence shown in FIG.1 (SEQ ID NO: 3), FIG. 3 (SEQ ID NO: 8), FIG. 5 (SEQ ID NO: 10), FIG. 7(SEQ ID NO: 12), FIG. 9 (SEQ ID NO: 14), FIG. 11 (SEQ ID NO: 16), FIG.13 (SEQ ID NO: 21), FIG. 15 (SEQ ID NO: 23), FIG. 17 (SEQ ID NO: 28),FIG. 19 (SEQ ID NO: 31), FIG. 21 (SEQ ID NO: 38), FIG. 23 (SEQ ID NO:40), FIG. 25 (SEQ ID NO: 51), FIG. 27 (SEQ ID NO: 53), FIG. 29 (SEQ IDNO: 55), FIG. 31 (SEQ ID NO: 57), FIG. 33 (SEQ ID NO: 62), FIG. 35 (SEQID NO: 64), FIG. 37 (SEQ ID NO: 72), FIG. 39 (SEQ ID NO: 77), FIG. 41(SEQ ID NO: 79), FIG. 43 (SEQ ID NO: 83), FIG. 45 (SEQ ID NO: 85), FIG.47 (SEQ ID NO: 90), FIG. 49 (SEQ ID NO: 98), FIG. 51 (SEQ ID NO: 103),FIG. 53 (SEQ ID NO: 105), FIG. 55 (SEQ ID NO: 107), FIG. 57 (SEQ ID NO:109), FIG. 59 (SEQ ID NO: 114), FIG. 61 (SEQ ID NO: 120), FIG. 63 (SEQID NO: 125), FIG. 65 (SEQ ID NO: 127), FIGS. 67A-B (SEQ ID NO: 129),FIG. 69 (SEQ ID NO: 131), FIG. 71 (SEQ ID NO: 136) and FIG. 73 (SEQ IDNO: 138).
 4. Isolated nucleic acid having at least 80% nucleic acidsequence identity to the full-length coding sequence of the DNAdeposited under any ATCC accession number shown in Table
 10. 5. A vectorcomprising the nucleic acid of any one of claims 1 to
 4. 6. The vectorof claim 5 operably linked to control sequences recognized by a hostcell transformed with the vector.
 7. A host cell comprising the vectorof claim
 5. 8. The host cell of claim 7, wherein said cell is a CHOcell.
 9. The host cell of claim 7, wherein said cell is an E. coli. 10.The host cell of claim 7, wherein said cell is a yeast cell.
 11. Aprocess for producing a PRO polypeptides comprising culturing the hostcell of claim 7 under conditions suitable for expression of said PROpolypeptide and recovering said PRO polypeptide from the cell culture.12. An isolated polypeptide having at least 80% amino acid sequenceidentity to an amino acid sequence selected from the group consisting ofthe amino acid sequence shown in FIG. 2 (SEQ ID NO: 4), FIG. 4 (SEQ IDNO: 9), FIG. 6 (SEQ ID NO: 11), FIG. 8 (SEQ ID NO: 13), FIG. 10 (SEQ IDNO: 15), FIG. 12 (SEQ ID NO: 17), FIG. 14 (SEQ ID NO: 22), FIG. 16 (SEQID NO: 24), FIG. 18 (SEQ ID NO: 29), FIG. 20 (SEQ ID NO: 32), FIG. 22(SEQ ID NO: 39), FIG. 24 (SEQ ID NO: 41), FIG. 26 (SEQ ID NO: 52), FIG.28 (SEQ ID NO: 54), FIG. 30 (SEQ ID NO: 56), FIG. 32 (SEQ ID NO: 58),FIG. 34 (SEQ ID NO: 63), FIG. 36 (SEQ ID NO: 65), FIG. 38 (SEQ ID NO:73), FIG. 40 (SEQ ID NO: 78), FIG. 42 (SEQ ID NO: 80), FIG. 44 (SEQ IDNO: 84), FIG. 46 (SEQ ID NO: 86), FIG. 48 (SEQ ID NO: 91), FIG. 50 (SEQID NO: 99), FIG. 52 (SEQ ID NO: 104), FIG. 54 (SEQ ID NO: 106), FIG. 56(SEQ ID NO: 108), FIG. 58 (SEQ ID NO: 110), FIG. 60 (SEQ ID NO: 115),FIG. 62 (SEQ ID NO: 121), FIG. 64 (SEQ ID NO: 126), FIG. 66 (SEQ ID NO:128), FIG. 68 (SEQ ID NO: 130), FIG. 70 (SEQ ID NO: 132), FIG. 72 (SEQID NO: 137) and FIG. 74 (SEQ ID NO: 139).
 13. An isolated polypeptidescoring at least 80% positives when compared to an amino acid sequenceselected from the group consisting of the amino acid sequence shown inFIG. 2 (SEQ ID NO: 4), FIG. 4 (SEQ ID NO: 9), FIG. 6 (SEQ ID NO: 11),FIG. 8 (SEQ ID NO: 13), FIG. 10 (SEQ ID NO: 15), FIG. 12 (SEQ ID NO:17), FIG. 14 (SEQ ID NO: 22), FIG. 16 (SEQ ID NO: 24), FIG. 18 (SEQ IDNO: 29), FIG. 20 (SEQ ID NO: 32), FIG. 22 (SEQ ID NO: 39), FIG. 24 (SEQID NO: 41), FIG. 26 (SEQ ID NO: 52), FIG. 28 (SEQ ID NO: 54), FIG. 30(SEQ ID NO: 56), FIG. 32 (SEQ ID NO: 58), FIG. 34 (SEQ ID NO: 63), FIG.36 (SEQ ID NO: 65), FIG. 38 (SEQ ID NO: 73), FIG. 40 (SEQ ID NO: 78),FIG. 42 (SEQ ID NO: 80), FIG. 44 (SEQ ID NO: 84), FIG. 46 (SEQ ID NO:86), FIG. 48 (SEQ ID NO: 91), FIG. 50 (SEQ ID NO: 99), FIG. 52 (SEQ IDNO: 104), FIG. 54 (SEQ ID NO: 106), FIG. 56 (SEQ ID NO: 108), FIG. 58(SEQ ID NO: 110), FIG. 60 (SEQ ID NO: 115), FIG. 62 (SEQ ID NO: 121),FIG. 64 (SEQ ID NO: 126), FIG. 66 (SEQ ID NO: 128), FIG. 68 (SEQ ID NO:130), FIG. 70 (SEQ ID NO: 132), FIG. 72 (SEQ ID NO: 137) and FIG. 74(SEQ ID NO: 139).
 14. An isolated polypeptide having at least 80% aminoacid sequence identity to an amino acid sequence encoded by thefull-length coding sequence of the DNA deposited under any ATCCaccession number shown in Table
 10. 15. A chimeric molecule comprising apolypeptide according to any one of claims 12 to 14 fused to aheterologous amino acid sequence.
 16. The chimeric molecule of claim 15,wherein said heterologous amino acid sequence is an epitope tagsequence.
 17. The chimeric molecule of claim 15, wherein saidheterologous amino acid sequence is a Fc region of an immunoglobulin.18. An antibody which specifically binds to a polypeptide according toany one of claims 12 to
 14. 19. The antibody of claim 18, wherein saidantibody is a monoclonal antibody, a humanized antibody or asingle-chain antibody.
 20. Isolated nucleic acid having at least 80%nucleic acid sequence identity to: (a) a nucleotide sequence encodingthe polypeptide shown in FIG. 2 (SEQ ID NO: 4), FIG. 4 (SEQ ID NO: 9),FIG. 6 (SEQ ID NO: 11), FIG. 8 (SEQ ID NO: 13), FIG. 10 (SEQ ID NO: 15),FIG. 12 (SEQ ID NO: 17), FIG. 14 (SEQ ID NO: 22), FIG. 16 (SEQ ID NO:24), FIG. 18 (SEQ ID NO: 29), FIG. 20 (SEQ ID NO: 32), FIG. 22 (SEQ IDNO: 39), FIG. 24 (SEQ ID NO: 41), FIG. 26 (SEQ ID NO: 52), FIG. 28 (SEQID NO: 54), FIG. 30 (SEQ ID NO: 56), FIG. 32 (SEQ ID NO: 58), FIG. 34(SEQ ID NO: 63), FIG. 36 (SEQ ID NO: 65), FIG. 38 (SEQ ID NO: 73), FIG.40 (SEQ ID NO: 78), FIG. 42 (SEQ ID NO: 80), FIG. 44 (SEQ ID NO: 84),FIG. 46 (SEQ ID NO: 86), FIG. 48 (SEQ ID NO: 91), FIG. 50 (SEQ ID NO:99), FIG. 52 (SEQ ID NO: 104), FIG. 54 (SEQ ID NO: 106), FIG. 56 (SEQ IDNO: 108), FIG. 58 (SEQ ID NO: 110), FIG. 60 (SEQ ID NO: 115), FIG. 62(SEQ ID NO: 121), FIG. 64 (SEQ ID NO: 126), FIG. 66 (SEQ ID NO: 128),FIG. 68 (SEQ ID NO: 130), FIG. 70 (SEQ ID NO: 132), FIG. 72 (SEQ ID NO:137) or FIG. 74 (SEQ ID NO: 139), lacking its associated signal peptide;(b) a nucleotide sequence encoding an extracellular domain of thepolypeptide shown in FIG. 2 (SEQ ID NO: 4), FIG. 4 (SEQ ID NO: 9), FIG.6 (SEQ ID NO: 11), FIG. 8 (SEQ ID NO: 13), FIG. 10 (SEQ ID NO: 15), FIG.12 (SEQ ID NO: 17), FIG. 14 (SEQ ID NO: 22), FIG. 16 (SEQ ID NO: 24),FIG. 18 (SEQ ID NO: 29), FIG. 20 (SEQ ID NO: 32), FIG. 22 (SEQ ID NO:39), FIG. 24 (SEQ ID NO: 41), FIG. 26 (SEQ ID NO: 52), FIG. 28 (SEQ IDNO: 54), FIG. 30 (SEQ ID NO: 56), FIG. 32 (SEQ ID NO: 58), FIG. 34 (SEQID NO: 63), FIG. 36 (SEQ ID NO: 65), FIG. 38 (SEQ ID NO: 73), FIG. 40(SEQ ID NO: 78), FIG. 42 (SEQ ID NO: 80), FIG. 44 (SEQ ID NO: 84), FIG.46 (SEQ ID NO: 86), FIG. 48 (SEQ ID NO: 91), FIG. 50 (SEQ ID NO: 99),FIG. 52 (SEQ ID NO: 104), FIG. 54 (SEQ ID NO: 106), FIG. 56 (SEQ ID NO:108), FIG. 58 (SEQ ID NO: 110), FIG. 60 (SEQ ID NO: 115), FIG. 62 (SEQID NO: 121), FIG. 64 (SEQ ID NO: 126), FIG. 66 (SEQ ID NO: 128), FIG. 68(SEQ ID NO: 130), FIG. 70 (SEQ ID NO: 132), FIG. 72 (SEQ ID NO: 137) orFIG. 74 (SEQ ID NO: 139), with its associated signal peptide; or (c) anucicotide sequence encoding an extracellular domain of the polypeptideshown in FIG. 2 (SEQ ID NO: 4), FIG. 4 (SEQ ID NO: 9), FIG. 6 (SEQ IDNO: 11), FIG. 8 (SEQ ID NO: 13), FIG. 10 (SEQ ID NO: 15), FIG. 12 (SEQID NO: 17), FIG. 14 (SEQ ID NO: 22), FIG. 16 (SEQ ID NO: 24), FIG. 18(SEQ ID NO: 29), FIG. 20 (SEQ ID NO: 32), FIG. 22 (SEQ ID NO: 39), FIG.24 (SEQ ID NO: 41), FIG. 26 (SEQ ID NO: 52), FIG. 28 (SEQ ID NO: 54),FIG. 30 (SEQ ID NO: 56), FIG. 32 (SEQ ID NO: 58), FIG. 34 (SEQ ID NO:63), FIG. 36 (SEQ ID NO: 65), FIG. 38 (SEQ ID NO: 73), FIG. 40 (SEQ IDNO: 78), FIG. 42 (SEQ ID NO: 80), FIG. 44 (SEQ ID NO: 84), FIG. 46 (SEQID NO: 86), FIG. 48 (SEQ ID NO: 91), FIG. 50 (SEQ ID NO: 99), FIG. 52(SEQ ID NO: 104), FIG. 54 (SEQ ID NO: 106), FIG. 56 (SEQ ID NO: 108),FIG. 58 (SEQ ID NO: 110), FIG. 60 (SEQ ID NO: 115), FIG. 62 (SEQ ID NO:121), FIG. 64 (SEQ ID NO: 126), FIG. 66 (SEQ ID NO: 128), FIG. 68 (SEQID NO: 130), FIG. 70 (SEQ ID NO: 132), FIG. 72 (SEQ ID NO: 137) or FIG.74 (SEQ ID NO: 139), lacking its associated signal peptide.
 21. Anisolated polypeptide having at least 80% amino acid, sequence identityto: (a) the polypeptide shown in FIG. 2 (SEQ ID NO: 4), FIG. 4 (SEQ IDNO: 9), FIG. 6 (SEQ ID NO: 11), FIG. 8 (SEQ ID NO: 13), FIG. 10 (SEQ IDNO: 15), FIG. 12 (SEQ ID NO: 17), FIG. 14 (SEQ ID NO: 22), FIG. 16 (SEQID NO: 24), FIG. 18 (SEQ ID NO: 29), FIG. 20 (SEQ ID NO: 32), FIG. 22(SEQ ID NO: 39), FIG. 24 (SEQ ID NO: 41), FIG. 26 (SEQ ID NO: 52), FIG.28 (SEQ ID NO: 54), FIG. 30 (SEQ ID NO: 56), FIG. 32 (SEQ ID NO: 58),FIG. 34 (SEQ ID NO: 63), FIG. 36 (SEQ ID NO: 65), FIG. 38 (SEQ ID NO:73), FIG. 40 (SEQ ID NO: 78), FIG. 42 (SEQ ID NO: 80), FIG. 44 (SEQ IDNO: 84), FIG. 46 (SEQ ID NO: 86), FIG. 48 (SEQ ID NO: 91), FIG. 50 (SEQID NO: 99), FIG. 52 (SEQ ID NO: 104), FIG. 54 (SEQ ID NO: 106), FIG. 56(SEQ ID NO: 108), FIG. 58 (SEQ ID NO: 110), FIG. 60 (SEQ ID NO: 115),FIG. 62 (SEQ ID NO: 121), FIG. 64 (SEQ ID NO: 126), FIG. 66 (SEQ ID NO:128), FIG. 68 (SEQ ID NO: 130), FIG. 70 (SEQ ID NO: 132), FIG. 72 (SEQID NO: 137) or FIG. 74 (SEQ ID NO: 139), lacking its associated signalpeptide; (b) an extracellular domain of the polypeptide shown in FIG. 2(SEQ ID NO: 4), FIG. 4 (SEQ ID NO: 9), FIG. 6 (SEQ ID NO: 11), FIG. 8(SEQ ID NO: 13), FIG. 10 (SEQ ID NO: 15), FIG. 12 (SEQ ID NO: 17), FIG.14 (SEQ ID NO: 22), FIG. 16 (SEQ ID NO: 24), FIG. 18 (SEQ ID NO: 29),FIG. 20 (SEQ ID NO: 32), FIG. 22 (SEQ ID NO: 39), FIG. 24 (SEQ ID NO:41), FIG. 26 (SEQ ID NO: 52), FIG. 28 (SEQ ID NO: 54), FIG. 30 (SEQ IDNO: 56), FIG. 32 (SEQ ID NO: 58), FIG. 34 (SEQ ID NO: 63), FIG. 36 (SEQID NO: 65), FIG. 38 (SEQ ID NO: 73), FIG. 40 (SEQ ID NO: 78), FIG. 42(SEQ ID NO: 80), FIG. 44 (SEQ ID NO: 84), FIG. 46 (SEQ ID NO: 86), FIG.48 (SEQ ID NO: 91), FIG. 50 (SEQ ID NO: 99), FIG. 52 (SEQ ID NO: 104),FIG. 54 (SEQ ID NO: 106), FIG. 56 (SEQ ID NO: 108), FIG. 58 (SEQ ID NO:110), FIG. 60 (SEQ ID NO: 115), FIG. 62 (SEQ ID NO: 121), FIG. 64 (SEQID NO: 126), FIG. 66 (SEQ ID NO: 128), FIG. 68 (SEQ ID NO: 130), FIG. 70(SEQ ID NO: 132), FIG. 72 (SEQ ID NO: 137) or FIG. 74 (SEQ ID NO: 139),with its associated signal peptide; or (c) an extracellular domain ofthe polypeptide shown in FIG. 2 (SEQ ID NO: 4), FIG. 4 (SEQ ID NO: 9),FIG. 6 (SEQ ID NO: 11), FIG. 8 (SEQ ID NO: 13), FIG. 10 (SEQ ID NO: 15),FIG. 12 (SEQ ID NO: 17), FIG. 14 (SEQ ID NO: 22), FIG. 16 (SEQ ID NO:24), FIG. 18 (SEQ ID NO: 29), FIG. 20 (SEQ ID NO: 32), FIG. 22 (SEQ IDNO: 39), FIG. 24 (SEQ ID NO: 41), FIG. 26 (SEQ ID NO: 52), FIG. 28 (SEQID NO: 54), FIG. 30 (SEQ ID NO: 56), FIG. 32 (SEQ ID NO: 58), FIG. 34(SEQ ID NO: 63), FIG. 36 (SEQ ID NO: 65), FIG. 38 (SEQ ID NO: 73), FIG.40 (SEQ ID NO: 78), FIG. 42 (SEQ ID NO: 80), FIG. 44 (SEQ ID NO: 84),FIG. 46 (SEQ ID NO: 86), FIG. 48 (SEQ ID NO: 91), FIG. 50 (SEQ ID NO:99), FIG. 52 (SEQ ID NO: 104), FIG. 54 (SEQ ID NO: 106), FIG. 56 (SEQ IDNO: 108), FIG. 58 (SEQ ID NO: 110), FIG. 60 (SEQ ID NO: 115), FIG. 62(SEQ ID NO: 121), FIG. 64 (SEQ ID NO: 126), FIG. 66 (SEQ ID NO: 128),FIG. 68 (SEQ ID NO: 130), FIG. 70 (SEQ ID NO: 132), FIG. 72 (SEQ ID NO:137) or FIG. 74 (SEQ ID NO: 139), lacking its associated signal peptide.22. A method of detecting a polypeptide designated as A, B, C, D, E, F,G, H, or I in a sample suspected of containing an A, B, C, D, E, F, G,H, or I polypeptide, said method comprising contacting said sample witha polypeptide designated herein as J, K, L, M, N, O, P, Q, R, S or T anddetermining the formation of a A/J, B/K, C/L, C/M, C/N, C/J, D/O, E/P,F/Q, G/R, H/S or I/T polypeptide conjugate in said sample, wherein theformation of said conjugate is indicative of the presence of an A, B, C,D, E, F, G, H, or I polypeptide in said sample and wherein A is a PRO533polypeptide, B is a PRO301 polypeptide, C is a PRO187 polypeptide, D isa PRO337 polypeptide, E is a PRO1411 polypeptide, F is a PRO10096polypeptide, G is a PRO246 polypeptide, H is a PRO6307 polypeptide, I isa PRO6003 polypeptide, J is an FGFR-4 polypeptide, K is a PRO301polypeptide, L is an FGFR-3 polypeptide, M is an FGFR-1 polypeptide, Nis an FGFR-2 polypeptide, O is a PRO6004 polypeptide, P is a PRO4356polypeptide, Q is a PRO2630 polypeptide, R is a PRO246 polypeptide, S isa PRO265 polypeptide and T is a PRO941 polypeptide.
 23. The methodaccording to claim 22, wherein said sample comprises cells suspected ofexpressing said A, B, C, D, E, F, G, H, or I polypeptide.
 24. The methodaccording to claim 22, wherein said J, K, L, M, N, O, P, Q, R, S or Tpolypeptide is labeled with a detectable label.
 25. The method accordingto claim 22, wherein said J, K, L, M, N, O, P, Q, R, S or T polypeptideis attached to a solid support.
 26. A method of detecting a polypeptidedesignated as J, K, L, M, N, O, P, Q, R, S or T in a sample suspected ofcontaining a J, K, L, M, N, O, P, Q, R, S or T polypeptide, said methodcomprising contacting said sample with a polypeptide designated hereinas A, B, C, D, E, F, G, H, or I and determining the formation of a A/J,B/K, C/L, C/M, C/N, C/J, D/O, E/P, F/Q, G/R, H/S or I/T polypeptideconjugate in said sample, wherein the formation of said conjugate isindicative of the presence of a J, K, L, M, N, O, P, Q, R, S or Tpolypeptide in said sample and wherein A is a PRO533 polypeptide, B is aPRO301 polypeptide, C is a PRO187 polypeptide, D is a PRO337polypeptide, E is a PRO1411 polypeptide, F is a PRO10096 polypeptide, Gis a PRO246 polypeptide, H is a PRO6307 polypeptide, I is a PRO6003polypeptide, J is an FGFR-4 polypeptide, K is a PRO301 polypeptide, L isan FGFR-3 polypeptide, M is an FGFR-1 polypeptide, N is an FGFR-2polypeptide, 0 is a PRO6004 polypeptide, P is a PRO4356 polypeptide, Qis a PRO2630 polypeptide, R is a PRO246 polypeptide, S is a PRO265polypeptide and T is a PRO941 polypeptide.
 27. The method according toclaim 26, wherein said sample comprises cells suspected of expressingsaid J, K, L, M, N, O, P, Q, R, S or T polypeptide.
 28. The methodaccording to claim 26, wherein said A, B, C, D, E, F, G, H, or Ipolypeptide is labeled with a detectable label.
 29. The method accordingto claim 26, wherein said A, B, C, D, E, F, G, H, or I polypeptide isattached to a solid support.
 30. A method of linking a bioactivemolecule to a cell expressing a polypeptide designated as A, B, C, D, E,F, G, H, I, said method comprising contacting said cell with apolypeptide designated as J, K, L, M, N, O, P, Q, R, S or T that isbound to said bioactive molecule and allowing said A, B, C, D, E, F, G,H, or I and said J, K, L, M, N, O, P, Q, R, S or T polypeptides to bindto one another, thereby linking said bioactive molecules to said cell,wherein A is a PRO533 polypeptide, B is a PRO301 polypeptide, C is aPRO187 polypeptide, D is a PRO337 polypeptide, E is a PRO1411polypeptide, F is a PRO10096 polypeptide, G is a PRO246 polypeptide, His a PRO6307 polypeptide, I is a PRO6003 polypeptide, J is an FGFR-4polypeptide, K is a PRO301 polypeptide, L is an FGFR-3 polypeptide, M isan FGFR-1 polypeptide, N is an FGFR-2 polypeptide, O is a PRO6004polypeptide, P is a PRO4356 polypeptide, Q is a PRO2630 polypeptide, Ris a PRO246 polypeptide, S is a PRO265 polypeptide and T is a PRO941polypeptide.
 31. The method according to claim 30, wherein saidbioactive molecule is a toxin, a radiolabel or an antibody.
 32. Themethod according to claim 30, wherein said bioactive molecule causes thedeath of said cell.
 33. A method of linking a bioactive molecule to acell expressing a polypeptide designated as J, K, L, M, N, O, P, Q, R, Sor T, said method comprising contacting said cell with a polypeptidedesignated as A, B, C, D, E, F, G, H, or I that is bound to saidbioactive molecule and allowing said A, B, C, D, E, F, G, H, or I andsaid J, K, L, M, N, O, P, Q, R, S or T polypeptides to bind to oneanother, thereby linking said bioactive molecules to said cell, whereinA is a PRO533 polypeptide, B is a PRO301 polypeptide, C is a PRO187polypeptide, D is a PRO337 polypeptide, E is a PRO1411 polypeptide, F isa PRO10096 polypeptide, G is a PRO246 polypeptide, H is a PRO6307polypeptide, I is a PRO6003 polypeptide, J is an FGFR-4 polypeptide, Kis a PRO301 polypeptide, L is an FGFR-3 polypeptide, M is an FGFR-1polypeptide, N is an FGFR-2 polypeptide, O is a PRO6004 polypeptide, Pis a PRO4356 polypeptide, Q is a PRO2630 polypeptide, R is a PRO246polypeptide, S is a PRO265 polypeptide and T is a PRO941 polypeptide.34. The method according to claim 33, wherein said bioactive molecule isa toxin, a radiolabel or an antibody.
 35. The method according to claim33, wherein said bioactive molecule causes the death of said cell.
 36. Amethod of modulating at least one biological activity of a cellexpressing a polypeptide designated as A, B, C, D, E, F, G, H, I, saidmethod comprising contacting said cell with a polypeptide designated asJ, K, L, M, N, O, P, Q, R, S or T or an anti-A, B, C, D, E, F, G, H, orI polypeptide antibody, whereby said J, K, L, M, N, O, P, Q, R, S or Tpolypeptide or anti-A, B, C, D, E, F, G, H, or I polypeptide antibodybinds to said A, B, C, D, E, F, G, H, or I polypeptide, therebymodulating at least one biological activity of said cell.
 37. The methodaccording to claim 36, wherein said cell is killed.
 38. A method ofmodulating at least one biological activity of a cell expressing apolypeptide designated as J, K, L, M, N, O, P, Q, R, S or T, said methodcomprising contacting said cell with a polypeptide designated as A, B,C, D, E, F, G, H, or I or an antic-J, K, L, M, N, O, P, Q, R, S or Tpolypeptide antibody, whereby said antic-J, K, L, M, N, O, P, Q, R, S orT polypeptide antibody or A, B, C, D, E, F, G, H, or I polypeptideantibody binds to said J, K, L, M, N, O, P, Q, R, S or T polypeptide,thereby modulating at least one biological activity of said cell. 39.The method according to claim 36, wherein said cell is killed.